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Sample records for b16 melanoma transplant

  1. Growth Inhibition of Re-Challenge B16 Melanoma Transplant by Conjugates of Melanogenesis Substrate and Magnetite Nanoparticles as the Basis for Developing Melanoma-Targeted Chemo-Thermo-Immunotherapy

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    Tomoaki Takada

    2009-01-01

    Full Text Available Melanogenesis substrate, N-propionyl-cysteaminylphenol (NPrCAP, is selectively incorporated into melanoma cells and inhibits their growth by producing cytotoxic free radicals. Magnetite nanoparticles also disintegrate cancer cells and generate heat shock protein (HSP upon exposure to an alternating magnetic field (AMF. This study tested if a chemo-thermo-immunotherapy (CTI therapy strategy can be developed for better management of melanoma by conjugating NPrCAP on the surface of magnetite nanoparticles (NPrCAP/M. We examined the feasibility of this approach in B16 mouse melanoma and evaluated the impact of exposure temperature, frequency, and interval on the inhibition of re-challenged melanoma growth. The therapeutic protocol against the primary transplanted tumor with or without AMF exposure once a day every other day for a total of three treatments not only inhibited the growth of the primary transplant but also prevented the growth of the secondary, re-challenge transplant. The heat-generated therapeutic effect was more significant at a temperature of 43∘C than either 41∘C or 46∘C. NPrCAP/M with AMF exposure, instead of control magnetite alone or without AMF exposure, resulted in the most significant growth inhibition of the re-challenge tumor and increased the life span of the mice. HSP70 production was greatest at 43∘C compared to that with 41∘C or 46∘C. CD+T cells were infiltrated at the site of the re-challenge melanoma transplant.

  2. Oxidative burst of neutrophils against melanoma B16-F10.

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    Zivkovic, Morana; Poljak-Blazi, Marija; Zarkovic, Kamelija; Mihaljevic, Danijela; Schaur, Rudolf Joerg; Zarkovic, Neven

    2007-02-01

    Intensive oxidative burst was determined by chemiluminescence of peripheral blood neutrophils of mice that were intramuscularly injected with melanoma B16-F10 and/or subcutaneously with Sephadex G-200. The neutrophils from papula developed at the site of Sephadex injection were cytotoxic for the B16-F10 cells in vitro. However, survival of Sephadex injected tumour-bearing mice was lower than of control animals bearing B16-F10, while their tumours grew faster and were less necrotic. Thus, it is likely that injection of Sephadex distracted the neutrophils from the tumour allowing faster progression of the tumour, indicating that neutrophils may have an important role in the host defence against malignant cells in the early stage of tumour development.

  3. Oxidative burst of neutrophils against melanoma B16-F10.

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    Zivkovic, Morana; Poljak-Blazi, Marija; Zarkovic, Kamelija; Mihaljevic, Danijela; Schaur, Rudolf Joerg; Zarkovic, Neven

    2007-02-01

    Intensive oxidative burst was determined by chemiluminescence of peripheral blood neutrophils of mice that were intramuscularly injected with melanoma B16-F10 and/or subcutaneously with Sephadex G-200. The neutrophils from papula developed at the site of Sephadex injection were cytotoxic for the B16-F10 cells in vitro. However, survival of Sephadex injected tumour-bearing mice was lower than of control animals bearing B16-F10, while their tumours grew faster and were less necrotic. Thus, it is likely that injection of Sephadex distracted the neutrophils from the tumour allowing faster progression of the tumour, indicating that neutrophils may have an important role in the host defence against malignant cells in the early stage of tumour development. PMID:16564616

  4. Effective adoptive transfer of haploidentical tumor-specific T cells in B 16-melanoma bearing mice

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    CUI Nai-peng; XIE Shao-jian; HAN Jin-sheng; MA Zhen-feng; CHEN Bao-ping; CAI Jian-hui

    2012-01-01

    Background Adoptive transfer of allogeneic tumor-specific T cells often results in severe graft-versus-host disease (GVHD).Here,we sought to maximize graft-versus-tumor and minimize GVHD by using haploidentical T cells in pre-irradiated B16-melanoma bearing mice.Methods C57BL/6 mice bearing B16-melanoma tumors were irradiated with 0,5,or 7 Gy total body irradiation (TBI),or 7 Gy TBI plus bone marrow transplantation.Tumor areas were measured every 3 days to assess the influence of irradiation treatment on tumor regression.B16-melanoma bearing mice were irradiated with 7 Gy TBI; sera and spleens were harvested at days 1,3,5,7,9,11,and 13 after irradiation.White blood cell levels were measured and transforming growth factor β1 (TGF-β1) and interleukin 10 (IL-10) levels in serum were detected using enzyme-linked immunosorbent assay (ELISA) kits.Real-time reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry were performed to test TGF-β1,IL-10 and Foxp3 mRNA levels and the proportion of CD4+CD25+Foxp3+ T-regulatory cells (Tregs) in spleens.B16-melanoma bearing C57BL/6 mice were irradiated with 7 Gy TBI followed by syngeneic (Syn1/Syn2) or haploidentical (Hap1/Hap2),dendritic cell-induced cytotoxic T lymphocytes (DC-CTLs) treatment,tumor areas and system GVHD were observed every 3 days.Mice were killed 21 days after the DC-CTLs adoptive transfer;histologic analyses of eyes,skin,liver,lungs,and intestine were then performed.Results Irradiation with 7 Gy TBI on the B16-melanoma-bearing mice did not influence tumor regression compared to the control group; however,it down-regulated the proportion of Tregs in spleens and the TGF-β1 and IL-10 levels in sera and spleens,suggesting inhibition of autoimmunity and intervention of tumor microenvironment.Adoptive transfer of haploidentical DC-CTLs significantly inhibited B16-melanoma growth.GVHD assessment and histology analysis showed no significant difference among the groups.Conclusion Adoptive transfer

  5. SimB16: modeling induced immune system response against B16-melanoma.

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    Francesco Pappalardo

    Full Text Available Immunological therapy of progressive tumors requires not only activation and expansion of tumor specific cytotoxic T lymphocytes (CTLs, but also an efficient effector phase including migration of CTLs in the tumor tissue followed by conjugation and killing of target cells. We report the application of an agent-based model to recapitulate both the effect of a specific immunotherapy strategy against B16-melanoma in mice and the tumor progression in a generic tissue section. A comparison of the in silico results with the in vivo experiments shows excellent agreement. We therefore use the model to predict a critical role for CD137 expression on tumor vessel endothelium for successful therapy and other mechanistic aspects. Experimental results are fully compatible with the model predictions. The biologically oriented in silico model derived in this work will be used to predict treatment failure or success in other pre-clinical conditions eventually leading new promising in vivo experiments.

  6. Stimulation of melanogenesis by scoparone in B16 melanoma cells

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    Jeong-yeh YANG; Jeung-hyun KOO; Young-gil SONG; Kang-beom KWON; Ju-hyung LEE; Hee-sook SOHN; Byung-hyun PARK; Eun-chung JHEE; Jin-woo PARK

    2006-01-01

    Aim: The effect of coumarin derivatives on melanogenesis was investigated in B16 murine melanoma cells. Methods: Melanin content and tyrosinase activity were analyzed spectrophotometrically. The expression of tyrosinase, tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein-2 (TRP-2) were measured either by reverse transcription-polymerase chain reaction (RT-PCR) or Western blot. Results: Among the coumarin derivatives studied, scoparone (6,7-dimethoxycoumarin) was the most potent; the 6- or 7-methoxy group was found to be essential for the stimulation of melanogenesis. The melanin content was greatly increased by scoparone in a dose-dependent manner; there was no cytotoxicity at the effective concentrations. Scoparone increased enzyme activity as well as protein and mRNA expression of tyrosinase. In addition, mRNA of TRP-1 and TRP-2 were also increased after treatment with scoparone. H-89, an inhibitor of protein kinase A (PKA), completely inhibited the scoparone-induced increase of melanogenesis and the tyrosinase protein. Conclusion: These results suggest that scoparone-induced stimulation of melanogenesis is likely to occur at the transcriptional level of melanogenesis-related enzymes through PKA signaling.

  7. The effects of Hominis Placenta extract on Melanin synthesis of B16 melanoma cells

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    Hyung-Sik Seo

    2006-01-01

    Objective : This research was carried out for the development of medicine for vitiligo treatment and focused on the effect of Hominis Placenta extract on Melanin synthesis of B16 melanoma cells. Methods : Acitivity of tyrosinase playing a vital role in synthesis of Melanin and the quantity of Melanin, which is the final product in cultured B16 melanoma cells, effects of Hominis Placenta extract were measured. Results : The results indicated that Hominis Placenta extract increased both t...

  8. 5-Azacytidine and 5-aza-2'-deoxycytidine behave as different antineoplastic agents in B16 melanoma.

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    Cortvrindt, R.; Bernheim, J.; Buyssens, N.; Roobol, K.

    1987-01-01

    The antiproliferative effects of 5-azacytidine (acaCyd) and 5-aza-2'-deoxycytidine (azadCyd) were studied in murine B16 melanoma and a series of B16 melanoma derived mutant strains with selective resistances to the respective drugs. The in vitro cytotoxicities of azaCyd and azadCyd on B16 wild type, expressed in terms of IC50 values, were found to be 5 microM and 0.2 microM, respectively. The in vitro cytotoxicity of both drugs was dependent on the duration of exposure. Uridine and cytidine w...

  9. The Inhibitory Effect of Endostatin and Doxycycline Administration on B16 Melanoma Angiogenesis and Cellular Proliferation

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    Lisha Qi; Shiwu Zhang; Danfang Zhang; Xiaojin Yin; Sen Wang; Baochun Sun

    2008-01-01

    OBJECTIVE To investigate the effect of endostatin and doxycycline on melanoma cellular proliferation and tumor angiogenesis.METHODS The effects of endostatin and doxvcycline were studied in mice transplanted with B16 melanoma cells.The mice were divided into 4 groups that were trea ted as follows:endostatin treatment(E group),doxycycline treatment(D group),endostatin plus doxycycline trearment(DE group),controls(C group)received no treatment.Following 9 days of treatment the tumor tissue was removed to compare the differences in the tumor necrotic rate and micro-vessel density (MVD)among the different groups.Immunohistochemical staining was conducted to detect the expression of proliferating cell nuclear antigen(PCNA)in the different groups.RESULTS The MVD of the 3 experimental groups was significantly less than the control group,(F=10.888,P<0.05),indicating that doxycycline and endostatin can inhibit tumor angiogenesis by decreasing the tumor blood supply.This effect results in inhibition of tumor cellular proliferation and promotion of tumor cell necrosis.The tumor cell necrotic ra te of the 3 experimental groups were all significantly higher than the C group(F=7.229,P<0.05)and the difference between the DE and C groups also was statistically significant.PCNA expression in all 3 experimental groups was statistically less than the C group(F=17.729,P<0.05).CONCLUSION The combined use of endostatin and doxycyCline in vivo can influence PCNA exDression and angiogenesis in melanoma,and significantly inhibit melanoma cellular proliferation.

  10. Effect of Chlorogenic Acid on Melanogenesis of B16 Melanoma Cells

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    Hao-Rong Li

    2014-08-01

    Full Text Available Chlorogenic acid (CGA, the ester formed between caffeic acid and l-quinic acid, is a widespread phenolic compound. It is part of the human diet, found in foods such as coffee, apples, pears, etc. CGA is also was widely used in cosmetics, but the effects of CGA on melanogenesis are unknown. In this study, we analyzed the effects of CGA on cell proliferation, melanin content and tyrosinase of B16 murine melanoma cells. Additionally, the enzymatic reactions of CGA in B16 melanoma cells lytic solution were detected by UV spectrophotometry. Results showed CGA at 30 and 60 μM significantly suppresses cell proliferation. 8-MOP at 100 μM significantly promotes cell proliferation, but CGA can counter this. Incubated for 24 h, CGA (500 μM improves melanogenesis while suppressing tyrosinase activity in B16 melanoma cells or 8-methoxypsoralen (8-MOP co-incubated B16 melanoma cells. After 12 h, B16 melanoma cell treatment with CGA leads to an increase in melanin accumulation, however, after 48 h there is a decrease in melanin production which correlates broadly with a decrease in tyrosinase activity. CGA incubated with lytic solution 24 h turned brown at 37 °C. The formation of new products (with a maximum absorption at 295 nm is associated with reduction of CGA (maximum absorption at 326 nm. Therefore, CGA has its two sidesroles in melanogenesis of B16 melanoma cells. CGA is a likely a substrate of melanin, but the metabolic product(s of CGA may suppress melanogenesis in B16 melanoma cells by inhibiting tyrosinase activity.

  11. Effects of total body irradiation on b16f10 melanoma-bearing mice%全身放射线照射对 B16 F10黑色素瘤小鼠的影响

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    王冰; 屈朋欢; 王艳华; 崔乃鹏; 蔡建辉; 陈保平

    2015-01-01

    目的:观察全身放射线照射( TBI)对B16F10黑色素瘤小鼠移植肿瘤生长及小鼠存活的影响。方法建立C57BL/6小鼠B16F10黑色素瘤移植肿瘤模型,采用不同剂量分别对小鼠进行TBI,观察小鼠移植肿瘤的生长和小鼠的存活情况;检测放疗后小鼠外周血白细胞水平。结果不同剂量TBI对各组小鼠肿瘤面积及存活率无影响(P均>0.05)。给予7 Gy TBI 10 d后,B16F10荷瘤小鼠外周血白细胞水平下降(P<0.05)。结论 TBI不影响B16 F10黑色素瘤小鼠移植肿瘤生长及荷瘤小鼠的生存;7 Gy TBI可改变荷瘤小鼠外周血白细胞水平,有利于肿瘤免疫治疗。%Objective To investigate effects of total body irradiation (TBI) on tumor growth and B16F10 melanoma-bearing mice survival.Methods C57BL/6 mice bearing B16-melanoma tumors were irradiated with 0, 5, or 7 Gy total body irradiation ( TBI) , or 7 Gy TBI pus bone marrow transplantation .Tumor areas were measured every 3 days to assess the influences of irradiation treatment on tumor regression .B16-melanoma bearing mice were irradiated with 7 Gy TBI and peripheral blood were harvested at days 1, 3, 5, 7, 9, 11 and 13 after irradiation to test WBC levels .Results TBI with variant dosage on the B 16-melanoma-bearing mice did not influence tumor regression compared with control group ( all P>0.05).WBC levels significantly decreased in the B16F10 melanoma-bearing mice on 10 d after 7 Gy TBI(P<0.05). Conclusion TBI dose do not influence tumor growth and survival of B 16F10 melanoma-bearing mice.Seven Gy TBI can alter WBC levels of peripheral bloods in B 16F10 melanoma-bearing mice, which helps to tumor immunotherapy .

  12. Fluvastatin increases tyrosinase synthesis induced by UVB irradiation of B16F10 melanoma cells.

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    Krzysztof Włodarski

    2010-02-01

    Full Text Available Statins are widely used to lower plasma concentrations of lipids, e.g. cholesterol. One of the main effects of statin treatment is inhibition of hydroxymethyl glutaryl-coenzyme A reductase. The role of fluvastatin, a frequently used statin, was examined in potential modulation of tyrosinase (key enzyme of melanogenesis synthesis. Levels of tyrosinase mRNA induced by UVB irradiation of B16F10 melanoma cell line were measured by real time PCR. Fluvastatin increases tyrosinase mRNA production induced by UVB irradiation in B16F10 melanoma cell line. Fluvastatin treatment may potentially influence melanin synthesis and protection against UV irradiation.

  13. Inhibitory effects of whisky congeners on melanogenesis in mouse B16 melanoma cells.

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    Ohguchi, Kenji; Koike, Minako; Suwa, Yoshihide; Koshimizu, Seiichi; Mizutani, Yuki; Nozawa, Yoshinori; Akao, Yukihiro

    2008-04-01

    We examined the effect of whisky congeners, substances other than ethanol in whisky, on melanogenesis in mouse B16 melanoma cells. Treatment with whisky congeners significantly blocked melanogenesis. Our results indicate that the inhibitory effects of whisky congeners on melanogenesis is due to direct inhibition of tyrosinase activity and to suppression of tyrosinase protein levels.

  14. Combination of amino acids reduces pigmentation in B16F0 melanoma cells.

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    Ishikawa, Masago; Kawase, Ichiro; Ishii, Fumio

    2007-04-01

    Amino acids, the building blocks of proteins, play significant roles in numerous physiological events in mammals. As the effects of amino acids on melanogenesis have yet to be demonstrated, the present study was conducted to identify whether amino acids, in particular alanine, glycine, isoleucine and leucine, influence melanogenesis in B16F0 melanoma cells. Glycine and L-isoleucine, but not D-isoleucine, reduced melanogenesis in a concentration-dependent manner without any morphological changes in B16F0 melanoma cells. L-Alanine and L-leucine, but not D-alanine and D-leucine, also reduced melanogenesis without any morphological changes in B16F0 melanoma cells. However these amino acids did not show a concentration-dependency. Combination of L-alanine and the other amino acids, particularly 4 amino acids combination, had an additive effect on the inhibition of melanogenesis compared with single treatment of L-alanine. None of the amino acids affected the activity of tyrosinase, a key enzyme in melanogenesis. These results suggest that L-alanine, glycine, L-isoleucine and L-leucine, but not the D-form amino acids, have a hypopigmenting effect in B16F0 melanoma cells, and that these effects are not due to the inhibition of tyrosinase activity. Combination of these 4 amino acids had the additive effect on hypopigmentation that was as similar as that of kojic acid. PMID:17409501

  15. 5-Azacytidine and 5-aza-2'-deoxycytidine behave as different antineoplastic agents in B16 melanoma.

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    Cortvrindt, R.; Bernheim, J.; Buyssens, N.; Roobol, K.

    1987-01-01

    The antiproliferative effects of 5-azacytidine (acaCyd) and 5-aza-2'-deoxycytidine (azadCyd) were studied in murine B16 melanoma and a series of B16 melanoma derived mutant strains with selective resistances to the respective drugs. The in vitro cytotoxicities of azaCyd and azadCyd on B16 wild type, expressed in terms of IC50 values, were found to be 5 microM and 0.2 microM, respectively. The in vitro cytotoxicity of both drugs was dependent on the duration of exposure. Uridine and cytidine were able to reverse the in vitro cytotoxicity of azaCyd, but not of azadCyd. Conversely, 2'-deoxycytidine was able to reverse the cytotoxic effect of azadCyd but not of azaCyd. Thymidine and 2'-deoxyuridine had no detectable effects on the in vitro cytotoxicity of either azaCyd or azadCyd. B16 melanoma mutant strains that were selected for resistance to azaCyd showed no cross-resistance to azadCyd, cytosine arabinoside or the fluorinated pyrimidine analogues FUrd, FCyd, FdUrd and FdCyd. Mutant strains that were selected for resistance to azadCyd showed no cross-resistance to azaCyd or fluorinated pyrimidine analogs, but only to cytosine arabinoside. The combined data suggest that azaCyd and azadCyd follow different routes of intracellular metabolic activation and exert their cytotoxic activity via different intracellular targets. PMID:2444244

  16. Sox10 controls migration of B16F10 melanoma cells through multiple regulatory target genes.

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    Ikjoo Seong

    Full Text Available It is believed that the inherent differentiation program of melanocytes during embryogenesis predisposes melanoma cells to high frequency of metastasis. Sox10, a transcription factor expressed in neural crest stem cells and a subset of progeny lineages, plays a key role in the development of melanocytes. We show that B16F10 melanoma cells transfected with siRNAs specific for Sox10 display reduced migratory activity which in turn indicated that a subset of transcriptional regulatory target genes of Sox10 is likely to be involved in migration and metastasis of melanoma cells. We carried out a microarray-based gene expression profiling using a Sox10-specific siRNA to identify relevant regulatory targets and found that multiple genes including melanocortin-1 receptor (Mc1r partake in the regulation of migration. We provide evidences that the effect of Sox10 on migration is mediated in large part by Mitf, a transcription factor downstream to Sox10. Among the mouse melanoma cell lines examined, however, only B16F10 showed robust down-regulation of Sox10 and inhibition of cell migration indicating that further dissection of dosage effects and/or cell line-specific regulatory networks is necessary. The involvement of Mc1r in migration was studied in detail in vivo using a murine metastasis model. Specifically, B16F10 melanoma cells treated with a specific siRNA showed reduced tendency in metastasizing to and colonizing the lung after being injected in the tail vein. These data reveal a cadre of novel regulators and mediators involved in migration and metastasis of melanoma cells that represents potential targets of therapeutic intervention.

  17. Electrochemotherapy by pulsed electromagnetic field treatment (PEMF) in mouse melanoma B16F10 in vivo

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    Kranjc Simona; Kranjc Matej; Scancar Janez; Jelenc Jure; Sersa Gregor; Miklavcic Damijan

    2016-01-01

    Introduction Pulsed electromagnetic field (PEMF) induces pulsed electric field, which presumably increases membrane permeabilization of the exposed cells, similar to the conventional electroporation. Thus, contactless PEMF could represent a promising approach for drug delivery. Materials and methods Noninvasive electroporation was performed by magnetic field pulse generator connected to an applicator consisting of round coil. Subcutaneous mouse B16F10 melanoma tumors were treated with intrave...

  18. [6]-Shogaol inhibits melanogenesis in B16 mouse melanoma cells through activation of the ERK pathway

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    Yao, Cheng; Oh, Jang-Hee; Oh, Inn Gyung; Park, Chi-Hyun; Chung, Jin Ho

    2012-01-01

    Aim: To investigate the effect of [6]-shogaol, an active ingredient in ginger, on melanogenesis and the underlying mechanisms. Methods: B16F10 mouse melanoma cells were tested. Cell viability was determined with the MTT assay. Melanin content and tyrosinase activity were analyzed with a spectrophotometer. The protein expression of tyrosinase and microphthalmia associated transcription factor (MITF), as well as phosphorylated or total ERK1/2 and Akt were measured using Western blot. Results: T...

  19. Melanogenesis Inhibitory Activity of Rhododendron Weyrichii in Mouse B16 Melanoma Cells.

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    Min-Jin Kim

    2016-08-01

    Full Text Available In this study, to evaluate the usefulness of Rhododendron weyrichii Maxim.as a whitening agent, the whitening effects of its extracts were investigated in alpha-melanocyte-stimulating hormone (α-MSH-induced B16F10 melanoma cells. No toxicity was noted in either B16F10 melanoma cells or HaCaT keratinocyte cells that were exposed to the hot water or 70% ethanol extracts of R. weyrichii Maxim. (RW-H and RW-E, respectively.Moreover, both the RW-H and RW-E extracts dose-dependently inhibited α-MSH-induced melanin production in B16F10 melanoma cells, with inhibitory effects of 52.5% and 51.6%, respectively, at a concentration of 200μg/mL. The RW-H and RW-E extracts also inhibitedintracellular tyrosinase activity in a dose-dependent fashion. Western blot analyses showed that the RW-H and RW-E extracts decreased tyrosinase, tyrosinase-relatedprotein-1, and tyrosinase-relatedprotein-2 expression.Additionally,we found that ρ-coumaric acid-containing RW-H and RW-E extracts could be used as hypopigmentation agentssince they suppress melanogenesis. Collectively, our results suggest that RW-H and RW-E extracts have the potential to serve as functional cosmetic agents, including whitening agents.

  20. Depigmenting Effect of Kojic Acid Esters in Hyperpigmented B16F1 Melanoma Cells

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    Ahmad Firdaus B. Lajis

    2012-01-01

    Full Text Available The depigmenting effect of kojic acid esters synthesized by the esterification of kojic acid using Rhizomucor miehei immobilized lipase was investigated in B16F1 melanoma cells. The depigmenting effect of kojic acid and kojic acid esters was evaluated by the inhibitory effect of melanin formation and tyrosinase activity on alpha-stimulating hormone- (α-MSH- induced melanin synthesis in B16F1 melanoma cells. The cellular tyrosinase inhibitory effect of kojic acid monooleate, kojic acid monolaurate, and kojic acid monopalmitate was found similar to kojic acid at nontoxic doses ranging from 1.95 to 62.5 μg/mL. However, kojic acid monopalmitate gave slightly higher inhibition to melanin formation compared to other inhibitors at doses ranging from 15.63 to 62.5 μg/mL. Kojic acid and kojic acid esters also show antioxidant activity that will enhance the depigmenting effect. The cytotoxicity of kojic acid esters in B16F1 melanoma cells was significantly lower than kojic acid at high doses, ranging from 125 and 500 μg/mL. Since kojic acid esters have lower cytotoxic effect than kojic acid, it is suggested that kojic acid esters can be used as alternatives for a safe skin whitening agent and potential depigmenting agents to treat hyperpigmentation.

  1. Superoxide dismutase activity as a function of culture aging of B-16 mouse melanoma cells

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    JOVANA B. SIMIC-KRSTIC

    2004-12-01

    Full Text Available The C3 clone of B-16 mouse melanoma was cultured for 1, 6 and 9 days and analysed. The changes which are not directly linked to melanogenesis in the B-16 / C3 cultures during their maturation were characterized. Early (1 day, confluent (6 days and old (9 days cell cultures are distinguished by their leucine aminopeptidase (LAP and a-naphthyl acetate esterase (ANAE isoenzyme patterns. Both quantitative and qualitative changes in LAP and ANAE isoenzyme can be observed during culture maturation. There is an increase in the activity of the enzyme copper, zinc-containing superoxide-dismutase (CuZn SOD. The increaase in the CuZn SOD enzyme activity might be related to B-16/C3 cell melanogenesis and / or to differentiation.

  2. DMEM enhances tyrosinase activity in B16 mouse melanoma cells and human melanocytes

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    Panpen Diawpanich

    2008-07-01

    Full Text Available Media components may affect the activities of cultured cells. In this study, tyrosinase activity was evaluated by using B16-F10 mouse melanoma cell lines (B16-F10 and primary human melanocytes cultured in different media. An optical density measurement and a L-dopa reaction assay were used as the determination of the tyrosinase activity. The study of B16-F10 found the optical density to be 2010, 2246 and 2961 in cells cultured in RPMI Medium 1640 (RPMI1640,Minimum Essential Medium (MEM and Dulbecco’s Modified Eagle Medium (DMEM, respectively. Moreover, compared to RPMI 1640 and MEM, DMEM showed the darkest color of melanin formation in culture media and in cells after the L-dopa reaction assay. Addition of kojic acid showed a significant inhibitory effect on tyrosinase activity in all media.Whereas MCDB153 showed no significant effect on human melanocytes, DMEM caused a dramatic increase in tyrosinase activity after 4 days of cultivation. Addition of kojic acid showed a significant tyrosinase inhibitory effect in DMEM only. Furthermore, an active ingredient in green tea, epigallocathechin gallate (EGCG could inhibit tyrosinase activity in both B16-F10 and human melanocytes cultured in DMEM. In summary, these results suggest that DMEM is a suitable medium that provides high detection sensitivity in a tyrosinase inhibition assay.

  3. Melanogenesis inhibitory activity of two generic drugs: cinnarizine and trazodone in mouse B16 melanoma cells.

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    Chang, Te-Sheng; Lin, Victor Chia-Hsiang

    2011-01-01

    More than 200 generic drugs were screened to identify the inhibitory activity on melanogenesis in mouse B16 melanoma cells. Cinnarizine and trazodone were identified as melanogenesis inhibitors. The inhibitory effects of the two drugs on cell survival, melanogenesis, and tyrosinase activity were investigated. The results showed that both cinnarizine and trazodone inhibited melanogenesis in B16 cells by a dose-dependent manner at the non-cytotoxic concentrations. Based on the results of the present study, seeking new melanogenesis inhibitors from generic drugs is an alternative approach to developing new depigmenting agents in cosmeceuticals. Moreover, cinnarizine and trazodone were proven to be good candidates as skin-whitening agents for treatment of skin hyperpigmentation. PMID:22272104

  4. Melanogenesis Inhibitory Activity of Two Generic Drugs: Cinnarizine and Trazodone in Mouse B16 Melanoma Cells

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    Te-Sheng Chang

    2011-12-01

    Full Text Available More than 200 generic drugs were screened to identify the inhibitory activity on melanogenesis in mouse B16 melanoma cells. Cinnarizine and trazodone were identified as melanogenesis inhibitors. The inhibitory effects of the two drugs on cell survival, melanogenesis, and tyrosinase activity were investigated. The results showed that both cinnarizine and trazodone inhibited melanogenesis in B16 cells by a dose-dependent manner at the non-cytotoxic concentrations. Based on the results of the present study, seeking new melanogenesis inhibitors from generic drugs is an alternative approach to developing new depigmenting agents in cosmeceuticals. Moreover, cinnarizine and trazodone were proven to be good candidates as skin-whitening agents for treatment of skin hyperpigmentation.

  5. The effect of antineoplastic drug NSC631570 on immunogenicity of B16 melanoma

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    Larysa M.Skivka

    2014-06-01

    Full Text Available Objectives: NSC631570 is a cytotoxic drug with the ability to be selectively accumulated in tumor tissue and activate apoptosis only in malignant cells and not in normal cells. Therapy with NSC631570 is accompanied by the stimulation of anticancer immune responses. It is known that cytotoxic anticancer drugs have additional effects on the immune system of tumor-bearing organism by increasing the immunogenic properties of tumor cells. This study aimed to investigate the immunogenicity of melanoma B16 after treatment with NSC631570. Methods: Two melanoma B16 sublines with different metastatic potentials were used. For in vivo growth cells were inoculated intravenously into C57BL/6 mice. The anticancer effect was calculated according to the growth inhibition index. Cell viability was determined by the MTT test. Cell apoptosis and necrosis were assessed by flow cytometry. HMGB1 expression, the serum level of cytokines and cytokine profile in tumor tissue were determined by ELISA. TAP1 and TAP2 expression was evaluated in RT-PCR and by Western blot. Results: Treatment of melanoma B16 cells with NSC631570 at apoptogenic concentrations induced tumor cell death accompanied by dose-dependent HMGB1 release in vitro. At the non-apoptogenic concentration the preparation caused an increase in TAP expression. The therapeutic efficacy of NSC631570 was also associated with strong release of HMGB1 in the serum of tumor-bearing mice and was more expressed in the case of the high-metastatic tumor variant. The therapeutic effect was accompanied by an increase of levels of Th1 cytokines in the serum and in tumor tissue of treated animals. Conclusion: In addition to the direct induction of tumor cell apoptosis, the preparation can increase the tumor immunogenicity. [J Exp Integr Med 2014; 4(2.000: 93-105

  6. Depigmenting mechanism of NSAIDs on B16F1 melanoma cells

    OpenAIRE

    Sato, Kazuomi; Takahashi, Hideki; Toriyama, Masaru

    2011-01-01

    The aim of the present work was to clarify the anti-melanogenic mechanism of non-steroidal anti-inflammatory drugs (NSAIDs). Mefenamic acid, diclofenac, and nimesulide were used in this study, and these drugs inhibit melanin synthesis in B16F1 melanoma cells. To elucidate the anti-melanogenic mechanism of NSAIDs, we performed western blotting analysis for melanogenic proteins, such as tyrosinase, TRP-1, and TRP-2. All NSAIDs used in this study inhibited tyrosinase protein level. Semi-quantita...

  7. Antineoplastic effects of Rhodiola crenulata treatment on B16-F10 melanoma.

    Science.gov (United States)

    Dudek, Maxine C; Wong, Kaitlyn E; Bassa, Lotfi M; Mora, Maria Carmen; Ser-Dolansky, Jennifer; Henneberry, Jean M; Crisi, Giovanna M; Arenas, Richard B; Schneider, Sallie S

    2015-12-01

    Melanoma is an aggressive form of skin cancer with limited treatment options for advanced stage disease. Early detection and wide surgical excision remain the initial mode of treatment for primary tumors thus preventing metastases and leading to improved prognosis. Through this work, we have evaluated the antineoplastic effects of Rhodiola crenulata (R. crenulata) root extracts on the B16-F10 melanoma cell line, both in vitro and in vivo. We observed that R. crenulata treatment resulted in increased cell death as well as a reduction in tumor cell proliferation and migration in vitro. Additionally, we observed that R. crenulata decreased the expression of integrin β1 and vimentin and increased the expression of E-cadherin. Further, in mice treated with a topical R. crenulata-based cream therapy, tumors were more likely to have a radial growth pattern, a reduction in mitotic activity, and an increase in tumor necrosis. We also observed that mice drinking water supplemented with R. crenulata displayed a reduction of metastatic foci in disseminated models of melanoma. Collectively, these findings suggest that R. crenulata exhibits striking antitumorigenic and antimetastatic properties and that this extract may harbor potential novel adjuvant therapy for the treatment of melanoma. PMID:26159852

  8. CCR4 is critically involved in effective antitumor immunity in mice bearing intradermal B16 melanoma.

    Science.gov (United States)

    Matsuo, Kazuhiko; Itoh, Tatsuki; Koyama, Atsushi; Imamura, Reira; Kawai, Shiori; Nishiwaki, Keiji; Oiso, Naoki; Kawada, Akira; Yoshie, Osamu; Nakayama, Takashi

    2016-08-01

    CCR4 is a major chemokine receptor expressed by Treg cells and Th17 cells. While Treg cells are known to suppress antitumor immunity, Th17 cells have recently been shown to enhance the induction of antitumor cytotoxic T lymphocytes. Here, CCR4-deficient mice displayed enhanced tumor growth upon intradermal inoculation of B16-F10 melanoma cells. In CCR4-deficient mice, while IFN-γ+CD8+ effector T cells were decreased in tumor sites, IFN-γ+CD8+ T cells and Th17 cells were decreased in regional lymph nodes. In wild-type mice, CD4+IL-17A+ cells, which were identified as CCR4+CD44+ memory Th17, were found to be clustered around dendritic cells expressing MDC/CCL22, a ligand for CCR4, in regional lymph nodes. Compound 22, a CCR4 antagonist, also enhanced tumor growth and decreased Th17 cells in regional lymph nodes in tumor-bearing mice treated with Dacarbazine. In contrast, CCR6 deficiency did not affect the tumor growth and the numbers of Th17 cells in regional lymph nodes. These findings indicate that CCR4 is critically involved in regional lymph node DC-Th17 cell interactions that are necessary for Th17 cell-mediated induction of antitumor CD8+ effector T cells in mice bearing B16 melanoma. PMID:27132989

  9. Melanogenesis inhibitory activity of sesquiterpenes from Canarium ovatum resin in mouse B16 melanoma cells.

    Science.gov (United States)

    Kikuchi, Takashi; Watanabe, Kensuke; Tochigi, Yuichi; Yamamoto, Ayako; Fukatsu, Makoto; Ezaki, Yoichiro; Tanaka, Reiko; Akihisa, Toshihiro

    2012-08-01

    Four known sesquiterpene alcohols, i.e., 1-4, ten triterpene alcohols, i.e., 5-14, and four triterpene acids, i.e., 15-18, were isolated from the MeOH extract of Canarium ovatum resin (elemi resin). Upon evaluation of the previously described compounds 1-18 on the melanogenesis in B16 melanoma cells induced with α-melanocyte-stimulating hormone (α-MSH), three sesquiterpene alcohols, i.e., cryptomeridiol (1), 4-epicryptomeridiol (2), and cadin-1(14)-ene-7α,11-diol (4), exhibited inhibitory effects with 27.4-34.1 and 39.0-56.9% reduction of melanin content at 50 and 100 μM, respectively, with no or very low toxicity to the cells (80.9-103.9% of cell viability at 100 μM). Western-blot analysis revealed that compounds 1 and 2 reduced the protein levels of MITF (=microphtalmia-associated transcription factor), tyrosinase, and TRP-2 (=tyrosine-related protein 2), mostly in a concentration-dependent manner, suggesting that these compounds exhibit melanogenesis inhibitory activity on α-MSH-stimulated B16 melanoma cells by, at least in part, inhibiting the expression of MITF, followed by decreasing the expression of tyrosinase and TRP-2. Three sesquiterpene alcohols, i.e., 1, 2, and 4, are, therefore, considered to be valuable as potential skin-whitening agents.

  10. Human placental lipid induces mitogenesis and melanogenesis in B16F10 melanoma cells

    Indian Academy of Sciences (India)

    Shampa Mallick; Samir Kumar Mandal; Ranjan Bhadra

    2002-06-01

    A hydroalcoholic extract of fresh term human placenta was found to be mitogenic as well as melanogenic on B16F10 mouse melanoma in an in vitro culture. The extract, a reservoir of a large number of bioactive molecules, was resolved to get the lipid fraction. Its activity was evaluated on B16F10 mouse melanoma by assessing the change in cellular morphology, growth and melanin induction. The lipid fraction, placental total lipid fraction (PTLF) tested in the study employed doses of 0.01 to 200 g/ml; optimum growth and melanization accompanied by morphological changes were recorded at 10 and 100 g/ml respectively. At intermediate doses growth and melanization were found to show a pattern of change over between growth and melanization and finally reached at an inverse relation at the respective optimal dose of response. Compared with defined sphingolipids, C2 ceramide and sphingosine-1-phosphate, the results were mostly corroborative. The duality of biological response of sphingolipids as reported in numerous studies was comparable for the PTLF suggesting that its active component is a sphingolipid and showing its use for pigment recovery in vitiligo.

  11. [6]-Shogaol inhibits melanogenesis in B16 mouse melanoma cells through activation of the ERK pathway

    Institute of Scientific and Technical Information of China (English)

    Cheng YAO; Jang-hee OH; Inn Gyung OH; Chi-hyun PARK; Jin Ho CHUNG

    2013-01-01

    Aim: To investigate the effect of [6]-shogaol,an active ingredient in ginger,on melanogenesis and the underlying mechanisms.Methods: B16F10 mouse melanoma cells were tested.Cell viability was determined with the MTT assay.Melanin content and tyrosinase activity were analyzed with a spectrophotometer.The protein expression of tyrosinase and microphthalmia associated transcription factor (MITF),as well as phosphorylated or total ERK1/2 and Akt were measured using Western blot.Results: Treatment of the cells with [6]-shogaol (1,5,10 μmol/L) reduced the melanin content in a concentration-dependent manner.[6]-Shogaol (5 and 10 μmol/L) significantly decreased the intracellular tyrosinase activity,and markedly suppressed the expression levels of tyrosinase and MITF proteins in the cells.Furthermore,[6]-shogaol (10 μmol/L) activated ERK,which was known to negatively regulate melanin synthesis in these cells.Pretreatment with the specific ERK pathway inhibitor PD98059 (20 μmol/L) greatly attenuated the inhibition of melanin synthesis by [6]-shogaol (10 μmol/L).Conclusion: The results demonstrate that [6]-shogaol inhibits melanogenesis in B16F10 mouse melanoma cells via activating the ERK pathway.

  12. The killing effect of 4-S-cysteaminylphenol, a newly synthesised melanin precursor, on B16 melanoma cell lines.

    OpenAIRE

    Yamada, I.; Seki, S.; Ito, S.; Suzuki, S.; Matsubara, O.; Kasuga, T.

    1991-01-01

    We have examined the killing effect of 4-S-cysteaminylphenol (4-S-CAP), a newly synthesised melanin precursor, on B16 melanoma cell lines possessing different melanin-producing activities and found it to be particularly effective in heavily melanised melanoma cells, but less so in moderately melanised melanoma cells, and having no effect on amelanotic melanoma cells and nonmelanoma cells. Thus, it was found that the killing effect of 4-S-CAP is highly dependent upon the synthesis of melanin a...

  13. Mechanism of low dose X-radiation influencing B16 melanoma blood-borne pulmonary metastases

    International Nuclear Information System (INIS)

    After whole body exposed to 7.5 cGy X-ray, the C57BL/6 mice were I. V. injected with 125IUdR labelled B16 melanoma cells, the proportion of surviving labelled cells retained in lungs was detected at 24 hours after injection, and found that the surviving cells in the lungs of irradiated mice were significantly decreased. Meanwhile, the immunofunctions were tested at 24 hours after irradiation, the results showed that the number of nucleate cells and NK cytotoxity in spleens, and the phagocytotic function of macrophages in vivo were increased obviously for the irradiated mice. The results suggested that the enhancement of NK cytotoxity and macrophage phagocytotic function might be one of the important reasons for low dose radiation accelerating the clearance of tumor cells from the lungs

  14. Atmospheric-pressure plasma jet characterization and applications on melanoma cancer treatment (B/16-F10)

    Energy Technology Data Exchange (ETDEWEB)

    Mashayekh, Shahriar [Physics Department, Shahid Beheshti University, G.C., Evin, 19839-63113 Tehran, Islamic Republic of Iran (Iran, Islamic Republic of); Rajaee, Hajar; Hassan, Zuhir M. [Imonology Department, Faculty of Medical Science, Tarbiat Modarres University, Tehran (Iran, Islamic Republic of); Akhlaghi, Morteza [Laser-Plasma Research Institute, Shahid Beheshti University, G.C., Evin, 19839-63113 Tehran, Islamic Republic of Iran (Iran, Islamic Republic of); Shokri, Babak [Physics Department and Laser-Plasma Research Institute, Shahid Beheshti University, G.C., Evin, 19839-63113 Tehran, Islamic Republic of Iran (Iran, Islamic Republic of)

    2015-09-15

    A new approach in medicine is the use of cold plasma for various applications such as sterilization blood coagulation and cancer cell treatment. In this paper, a pin-to-hole plasma jet for biological applications has been designed and manufactured and characterized. The characterization includes power consumption via Lissajous method, thermal behavior of atmospheric-pressure plasma jet by using Infra-red camera as a novel method and using Speicair software to determine vibrational and transitional temperatures, and optical emission spectroscopy to determine the generated species. Treatment of Melanoma cancer cells (B16/F10) was also implemented, and tetrazolium salt dye (MTT assay) and flow cytometry were used to evaluate viability. Effect of ultraviolet photons on cancerous cells was also observed using an MgF{sub 2} crystal with MTT assay. Finally, in-vivo studies on C57 type mice were also done in order to have a better understanding of the effects in real conditions.

  15. Atmospheric-pressure plasma jet characterization and applications on melanoma cancer treatment (B/16-F10)

    Science.gov (United States)

    Mashayekh, Shahriar; Rajaee, Hajar; Akhlaghi, Morteza; Shokri, Babak; Hassan, Zuhir M.

    2015-09-01

    A new approach in medicine is the use of cold plasma for various applications such as sterilization blood coagulation and cancer cell treatment. In this paper, a pin-to-hole plasma jet for biological applications has been designed and manufactured and characterized. The characterization includes power consumption via Lissajous method, thermal behavior of atmospheric-pressure plasma jet by using Infra-red camera as a novel method and using Speicair software to determine vibrational and transitional temperatures, and optical emission spectroscopy to determine the generated species. Treatment of Melanoma cancer cells (B16/F10) was also implemented, and tetrazolium salt dye (MTT assay) and flow cytometry were used to evaluate viability. Effect of ultraviolet photons on cancerous cells was also observed using an MgF2 crystal with MTT assay. Finally, in-vivo studies on C57 type mice were also done in order to have a better understanding of the effects in real conditions.

  16. Atmospheric-pressure plasma jet characterization and applications on melanoma cancer treatment (B/16-F10)

    International Nuclear Information System (INIS)

    A new approach in medicine is the use of cold plasma for various applications such as sterilization blood coagulation and cancer cell treatment. In this paper, a pin-to-hole plasma jet for biological applications has been designed and manufactured and characterized. The characterization includes power consumption via Lissajous method, thermal behavior of atmospheric-pressure plasma jet by using Infra-red camera as a novel method and using Speicair software to determine vibrational and transitional temperatures, and optical emission spectroscopy to determine the generated species. Treatment of Melanoma cancer cells (B16/F10) was also implemented, and tetrazolium salt dye (MTT assay) and flow cytometry were used to evaluate viability. Effect of ultraviolet photons on cancerous cells was also observed using an MgF2 crystal with MTT assay. Finally, in-vivo studies on C57 type mice were also done in order to have a better understanding of the effects in real conditions

  17. Cytosolic DNA Sensor Upregulation Accompanies DNA Electrotransfer in B16.F10 Melanoma Cells.

    Science.gov (United States)

    Znidar, Katarina; Bosnjak, Masa; Cemazar, Maja; Heller, Loree C

    2016-06-07

    In several preclinical tumor models, antitumor effects occur after intratumoral electroporation, also known as electrotransfer, of plasmid DNA devoid of a therapeutic gene. In mouse melanomas, these effects are preceded by significant elevation of several proinflammatory cytokines. These observations implicate the binding and activation of intracellular DNA-specific pattern recognition receptors or DNA sensors in response to DNA electrotransfer. In tumors, IFNβ mRNA and protein levels significantly increased. The mRNAs of several DNA sensors were detected, and DAI, DDX60, and p204 tended to be upregulated. These effects were accompanied with reduced tumor growth and increased tumor necrosis. In B16.F10 cells in culture, IFNβ mRNA and protein levels were significantly upregulated. The mRNAs for several DNA sensors were present in these cells; DNA-dependent activator of interferon regulatory factor (DAI), DEAD (Asp-Glu-Ala-Asp) box polypeptide 60 (DDX60), and p204 were significantly upregulated while DDX60 protein levels were coordinately upregulated. Upregulation of DNA sensors in tumors could be masked by the lower transfection efficiency compared to in vitro or to dilution by other tumor cell types. Mirroring the observation of tumor necrosis, cells underwent a significant DNA concentration-dependent decrease in proliferation and survival. Taken together, these results indicate that DNA electrotransfer may cause the upregulation of several intracellular DNA sensors in B16.F10 cells, inducing effects in vitro and potentially in vivo.

  18. Cytosolic DNA Sensor Upregulation Accompanies DNA Electrotransfer in B16.F10 Melanoma Cells

    Science.gov (United States)

    Znidar, Katarina; Bosnjak, Masa; Cemazar, Maja; Heller, Loree C.

    2016-01-01

    In several preclinical tumor models, antitumor effects occur after intratumoral electroporation, also known as electrotransfer, of plasmid DNA devoid of a therapeutic gene. In mouse melanomas, these effects are preceded by significant elevation of several proinflammatory cytokines. These observations implicate the binding and activation of intracellular DNA-specific pattern recognition receptors or DNA sensors in response to DNA electrotransfer. In tumors, IFNβ mRNA and protein levels significantly increased. The mRNAs of several DNA sensors were detected, and DAI, DDX60, and p204 tended to be upregulated. These effects were accompanied with reduced tumor growth and increased tumor necrosis. In B16.F10 cells in culture, IFNβ mRNA and protein levels were significantly upregulated. The mRNAs for several DNA sensors were present in these cells; DNA-dependent activator of interferon regulatory factor (DAI), DEAD (Asp-Glu-Ala-Asp) box polypeptide 60 (DDX60), and p204 were significantly upregulated while DDX60 protein levels were coordinately upregulated. Upregulation of DNA sensors in tumors could be masked by the lower transfection efficiency compared to in vitro or to dilution by other tumor cell types. Mirroring the observation of tumor necrosis, cells underwent a significant DNA concentration-dependent decrease in proliferation and survival. Taken together, these results indicate that DNA electrotransfer may cause the upregulation of several intracellular DNA sensors in B16.F10 cells, inducing effects in vitro and potentially in vivo. PMID:27271988

  19. Radiation Changes the Metabolic Profiling of Melanoma Cell Line B16

    Science.gov (United States)

    Yu, Yating; Liang, Wei; Zheng, Qinghui; Huang, Xianing; Huang, Yong; Lu, Xiaoling; Zhao, Yongxiang

    2016-01-01

    Radiation therapy can be an effective way to kill cancer cells using ionizing radiation, but some tumors are resistant to radiation therapy and the underlying mechanism still remains elusive. It is therefore necessary to establish an appropriate working model to study and monitor radiation-mediated cancer therapy. In response to cellular stress, the metabolome is the integrated profiling of changes in all metabolites in cells, which can be used to investigate radiation tolerance mechanisms and identify targets for cancer radiation sensibilization. In this study, using 1H nuclear magnetic resonance for untargeted metabolic profiling in radiation-tolerant mouse melanoma cell line B16, we comprehensively investigated changes in metabolites and metabolic network in B16 cells in response to radiation. Principal component analysis and partial least squares discriminant analysis indicated the difference in cellular metabolites between the untreated cells and X-ray radiated cells. In radiated cells, the content of alanine, glutamate, glycine and choline was increased, while the content of leucine, lactate, creatine and creatine phosphate was decreased. Enrichment analysis of metabolic pathway showed that the changes in metabolites were related to multiple metabolic pathways including the metabolism of glycine, arginine, taurine, glycolysis, and gluconeogenesis. Taken together, with cellular metabolome study followed by bioinformatic analysis to profile specific metabolic pathways in response to radiation, we deepened our understanding of radiation-resistant mechanisms and radiation sensibilization in cancer, which may further provide a theoretical and practical basis for personalized cancer therapy. PMID:27631970

  20. Comparison of the inhibitory effects of vitamin E analogues on melanogenesis in mouse B16 melanoma cells

    OpenAIRE

    Kamei, Yuto; Otsuka, Yuri; Abe, Kouichi

    2009-01-01

    The effect of eight vitamin E analogues (d-α-, dl-α-, d-β-, d-γ-, and d-δ-tocopherols, d-α- and dl-α-tocopheryl acetates) and 2,2,5,7,8-pentamethyl-6-hydroxychroman (PMC) on melanogenesis were compared in mouse B16 melanoma cells. D-β-tocopherol at 250 μg ml−1 inhibited not only 28% of melanin synthesis in B16 cells, but also 34% of the tyrosinase activity, a very important cascade enzyme involved in the synthesis of melanin in melanoma cells. D-γ-tocopherol also strongly inhibited up to 39% ...

  1. Electroporation transiently decreases GJB2 (connexin 26) expression in B16/BL6 melanoma cell line.

    Science.gov (United States)

    Rangel, Marcelo Monte Mór; Chaible, Lucas Martins; Nagamine, Marcia Kazumi; Mennecier, Gregory; Cogliati, Bruno; de Oliveira, Krishna Duro; Fukumasu, Heidge; Sinhorini, Idércio Luiz; Mir, Lluis Maria; Dagli, Maria Lúcia Zaidan

    2015-02-01

    Connexins are proteins that form gap junctions. Perturbations in the cell membrane reportedly promote changes in the expression profile of connexins. Electroporation promotes destabilization by applying electrical pulses, and this procedure is used in electrochemotherapy and gene therapy, among others. This in vitro work aimed to study the interference of electroporation on the expression profile of GJB2 (Cx26 gene) and Connexin 26 in melanoma cell line B16/BL6. The techniques of immunocytochemistry, Western blot, and real-time PCR were used. After electroporation, cells showed a transient decrease in GJB2 mRNA. The immunostaining of Cx26 showed no noticeable change after electroporation at different time points. However, Western blot showed a significant reduction in Cx26 30 min after electroporation. Our results showed that electroporation interferes transiently in the expression of Connexin 26 in melanoma and are consistent with the idea that electroporation is a process of intense stress that promotes cell homeostatic imbalance and results in disruption of cell physiological processes such as transcription and translation.

  2. An ester extract of Cochinchina momordica seeds induces differentiation of melanoma B16 F1 cells via MAPKs signaling.

    Science.gov (United States)

    Zhao, Lian-Mei; Han, Li-Na; Ren, Feng-Zhi; Chen, Shu-Hong; Liu, Li-Hua; Wang, Ming-Xia; Sang, Mei-Xiang; Shan, Bao-En

    2012-01-01

    Cochinchina momordica seeds (CMS) have been widely used due to antitumor activity by Mongolian tribes of China. However, the details of the underlying mechanisms remain unknown. In the present study, we found that an EtOAc (ethyl ester) extract of CMS (CMSEE) induced differentiation and caused growth inhibition of melanoma B16 F1 cells. CMSEE at the concentration of 5-200 μg/ml exhibited strongest anti-proliferative effects on B16 F1 cells among other CMS fractions (water or petroleum ether). Moreover, CMSEE induced melanoma B16 F1 cell differentiation, characterized by dendrite-like outgrowth, increasing melanogenesis production, as well as enhancing tyrosinase activity. Western blot analysis showed that sustained phosphorylation of p38 MAP accompanied by decrease in ERK1/2 and JNK dephosphorylation were involved in CMSEE-induced B16 F1 cell differentiation. Notably, 6 compounds that were isolated and identified may be responsible for inducing differentiation of CMSEE. These results indicated that CMSEE contributes to the differentiation of B16 F1 cells through modulating MAPKs activity, which may throw some light on the development of potentially therapeutic strategies for melanoma treatment. PMID:23098473

  3. B16 melanoma tumor growth is delayed in mice in an age-dependent manner

    Directory of Open Access Journals (Sweden)

    Christina Pettan-Brewer

    2012-08-01

    Full Text Available A major risk factor for cancer is increasing age, which suggests that syngeneic tumor implants in old mice would grow more rapidly. However, various reports have suggested that old mice are not as permissive to implanted tumor cells as young mice. In order to determine and characterize the age-related response to B16 melanoma, we implanted 5×105 tumor cells into 8, 16, 24, and 32-month-old male C57BL/6 (B6 and C57BL/6×BALB/c F1 (CB6 F1 mice subcutaneously in the inguinal and axillary spaces, or intradermally in the lateral flank. Results showed decreased tumor volume with increasing age, which varied according to mouse genetic background and the implanted site. The B6 strain showed robust tumor growth at 8 months of age at the inguinal implantation site, with an average tumor volume of 1341.25 mm3. The 16, 24, and 32-month age groups showed a decrease in tumor growth with tumor volumes of 563.69, 481.02, and 264.55 mm3, respectively (p≤0.001. The axillary implantation site was less permissive in 8-month-old B6 mice with an average tumor volume of 761.52 mm3. The 24- and 32-month age groups showed a similar decrease in tumor growth with tumor volumes of 440 and 178.19 mm3, respectively (p≤0.01. The CB6F1 strain was not as tumor permissive at 8 months of age as B6 mice with average tumor volumes of 446.96 and 426.91 mm3 for the inguinal and axillary sites, respectively. There was a decrease in tumor growth at 24 months of age at both inguinal and axillary sites with an average tumor volume of 271.02 and 249.12 mm3, respectively (p≤0.05. The strain dependence was not apparent in 8-month-old mice injected intradermally with B16 melanoma cells, with average tumor volumes of 736.82 and 842.85 mm3 for B6 and CB6 F1, respectively. However, a strain difference was seen in 32-month-old B6 mice with an average decrease in tumor volume of 250.83 mm3 (p≤0.01. In contrast, tumor growth significantly decreased earlier in CB6 F1 mice with average

  4. Cutaneous melanoma in solid organ transplant patients.

    Science.gov (United States)

    Russo, I; Piaserico, S; Belloni-Fortina, A; Alaibac, M

    2014-08-01

    Solid organ transplant patients are at greatly increased risk of developing a wide variety of skin cancers, particularly epithelial skin cancers. On the other hand, it is well known that an intact immune system limits the development of benign melanocytic lesions. The eruptive nevi phenomenon, which we can observe in solid organ transplant recipients, is indicative of the relationship between melanocyte proliferation and immune system. Regression of melanocytic nevi after restoration of complete immune responsiveness is a further clinical example the role of immunosurveillance on melanocyte proliferation. However, melanoma incidence in organ transplant recipients appears only 2-3 folds higher than in general population. To this regard, organ transplant recipients who develop de novo melanomas thicker than 2mm seem to have a significantly worse outcome with a greatly increased risk of dying of metastatic melanoma, whereas those who develop a ≤2 mm thickness melanoma seem to have a prognosis similar to that of the general population. Furthermore, there is no evidence supporting an increased risk of melanoma recurrences after transplant in patients with a history of low-risk melanoma. Melanoma is also one of the most frequent and lethal donor-derived malignancies suggesting that a history of invasive melanoma should be considered an absolute contraindication to donation. The aim of this review is to investigate the relationship between immunosuppression and melanoma and to discuss its clinical implications for the management of transplant-associated melanoma. PMID:25068225

  5. Anti-Melanogenic Property of Geoditin A in Murine B16 Melanoma Cells

    Directory of Open Access Journals (Sweden)

    Chun-Tao Che

    2012-02-01

    Full Text Available Geoditin A, an isomalabaricane triterpene isolated from marine sponge Geodia japonica, has been demonstrated to induce apoptosis in leukemia HL60 cells and human colon HT29 cancer cells through an oxidative stress, a process also interfering with normal melanogenesis in pigment cells. Treatment of murine melanoma B16 cells with geoditin A decreased expression of melanogenic proteins and cell melanogenesis which was aggravated with adenylate cyclase inhibitor SQ22536, indicating melanogenic inhibition was mediated through a cAMP-dependent signaling pathway. Immunofluorescence microscopy and glycosylation studies revealed abnormal glycosylation patterns of melanogenic proteins (tyrosinase and tyrosinase-related protein 1, and a co-localization of tyrosinase with calnexin (CNX and lysosome-associated membrane protein 1 (LAMP-1, implicating a post-translational modification in the ER and a degradation of tyrosinase in the lysosome. Taken together, potent anti-melanogenic property and the relatively low cytotoxicity of geoditin A have demonstrated its therapeutic potential as a skin lightening agent.

  6. Donor Transmission of Melanoma Following Renal Transplant

    Directory of Open Access Journals (Sweden)

    Kathryn T. Chen

    2012-01-01

    Full Text Available Donor transmission of melanoma is one of the more common and lethal of recipient malignancies, often presenting with systemic disease. Although some patients may receive durable remission of melanoma following explantation of the allograft and withdrawal of immunosuppression, donor transmission of melanoma is fatal in most patients. Here we present a case of a 44-year-old male who developed metastatic melanoma following renal transplant.

  7. Donor transmission of melanoma following renal transplant.

    Science.gov (United States)

    Chen, Kathryn T; Olszanski, Anthony; Farma, Jeffrey M

    2012-01-01

    Donor transmission of melanoma is one of the more common and lethal of recipient malignancies, often presenting with systemic disease. Although some patients may receive durable remission of melanoma following explantation of the allograft and withdrawal of immunosuppression, donor transmission of melanoma is fatal in most patients. Here we present a case of a 44-year-old male who developed metastatic melanoma following renal transplant.

  8. Alternol inhibits the proliferation and induces the differentiation of the mouse melanoma B16F0 cell line.

    Science.gov (United States)

    Wang, Caixia; Xu, Wenjuan; Hao, Wenjin; Wang, Bingsheng; Zheng, Qiusheng

    2016-08-01

    High malignant potential and low susceptibility to treatment are characteristics of malignant melanoma. Alternol, a novel compound purified from microbial fermentation products obtained from the bark of the yew tree, exhibits a variety of antitumor activities. Based on these findings, the aim of the present study was to extend the knowledge on the antineoplastic effect of alternol in the mouse melanoma B16F0 cell line. Alternol significantly inhibited the proliferation and colony formation of B16F0 cells in a dose-dependent manner as detected by MTT and soft agar colony formation assays. NaOH alkaline lysis and oxidation of Dopa indicated that alternol enhanced the melanin content and tyrosinase activity of the B16F0 cells and results also showed a dose‑response relationship. Morphologic changes accompanied by extended dendrites were discovered in the B16F0 cells after treatment with alternol. Furthermore, the mRNA levels of tyrosinase, Trp1 and Trp2 were increased by alternol. Our results confirmed that alternol possesses marked antineoplastic properties against melanoma cells, indicating that this microbial fermentation product is a promising agent for the differentiation therapy of cancer. The inhibition of cell proliferation and colony formation by alternol was associated with both cytotoxicity and induction of differentiation.

  9. Nanobiotechnological Nanocapsules Containing Polyhemoglobin-Tyrosinase: Effects on Murine B16F10 Melanoma Cell Proliferation and Attachment

    Directory of Open Access Journals (Sweden)

    Yun Wang

    2012-01-01

    Full Text Available We have reported previously that daily intravenous infusions of a soluble nanobiotechnological complex, polyhemoglobin-tyrosinase [polyHb-Tyr], can suppress the growth of murine B16F10 melanoma in a mouse model. In order to avoid the need for daily intravenous injections, we have now extended this further as follows. We have prepared two types of biodegradable nanocapsules containing [polyHb-Tyr]. One type is to increase the circulation time and decrease the frequency of injection and is based on polyethyleneglycol-polylactic acid (PEG-PLA nanocapsules containing [polyHb-Tyr]. The other type is to allow for intratumoural or local injection and is based on polylactic acid (PLA nanocapsules containing [polyHb-Tyr]. Cell culture studies show that it can inhibit the proliferation of murine B16F10 melanoma cells in the “proliferation model”. It can also inhibit the attachment of murine B16F10 melanoma cells in the “attachment model.” This could be due to the action of tyrosinase on the depletion of tyrosine or the toxic effect of tyrosine metabolites. The other component, polyhemoglobin (polyHb, plays a smaller role in nanocapsules containing [polyHb-Tyr], and this is most likely by its depletion of nitric oxide needed for melanoma cell growth.

  10. Anti-metastatic effects of the sulfated polysaccharide ascophyllan isolated from Ascophyllum nodosum on B16 melanoma.

    Science.gov (United States)

    Abu, Ryogo; Jiang, Zedong; Ueno, Mikinori; Isaka, Shogo; Nakazono, Satoru; Okimura, Takasi; Cho, Kichul; Yamaguchi, Kenichi; Kim, Daekyung; Oda, Tatsuya

    2015-03-20

    We previously found that ascophyllan, a sulfated polysaccharide isolated from brown seaweed Ascophyllum nodosum, exhibited antitumor activity in sarcoma-180 tumor-bearing mice. In this study, we found that ascophyllan inhibited the migration and adhesion of B16 melanoma cells by reducing the expression of N-cadherin and enhancing the expression of E-cadherin in a concentration-dependent manner. Transwell invasion assay revealed that ascophyllan suppressed the invasion ability of B16 cells. It also inhibited the expression of matrix metalloprotease-9 (MMP-9) mRNA and the secretion of MMP-9 protein in B16 cells, a process that may involve the extracellular signal-regulated kinase (ERK) signaling pathway. Furthermore, ascophyllan administered intraperitoneally at 25 mg/kg showed anti-metastatic activity in a mouse model of metastasis induced by intravenous injection of B16 cells, and the number of lung surface metastatic nodules in ascophyllan-treated mice was significantly reduced compared to that in the untreated control mice. Since splenic natural killer cell activity enhanced in the mice injected with ascophyllan intraperitoneally, we suggest that ascophyllan may exhibit in vivo anti-metastatic activity on B16 melanoma cells through activation of the host immune system in addition to a direct action on cancer cells. PMID:25623538

  11. 阿维A对鼠B16黑素瘤的增殖抑制及诱导分化作用%Acitretin inhibits the growth and induces the differentiation of mouse B16 melanoma

    Institute of Scientific and Technical Information of China (English)

    丁政云; 杨阳

    2009-01-01

    Objective To study the inhibition of growth and induction of differentiation of mouse B16 melanoma by acitretin and their mechanism.Methods Animal models of B16 melanoma were established by subcutaneously inoculation of cultured B16 cells into the right axilla of mice.All mice were divided into 5 groups,negative control group treated with peanut oil,low-dose acitretin group treated with acitretin 10 mg per kilogram of body weight per day,high-dose acitretin group treated with 20 mg per kilogram body weight per day,cisplatin group treated with cisplatin 10 mg per kilogram body weight,combination group treated with acitretin 20 mg per kilogram body weight per day plus cisplatin 10 mg per kilogram body weight.Acitretin was given daily via intragastric administration.and cisplatin was given with an interval of 7 days,from day 2 till day 22 after the inoculation.The growth of transplanted tumor was measured with an interval of 3 days.After drug withdrawal,mice were killed,transplanted tumors were obtained for the measurement of tumor weight,pathological examination and immunohistochemical staining for survivin,Fas and vascular endothelial growth factor(VEGF).Results Acitretin could significantly inhibit the growth of B16 melanoma,the average weight and volume of transplanted tumor in the treated groups were significantly lower than those in the negative control group(all P<0.01).Pathological examination revealed that in the control group,tumor cells showed typical heteromorphism,and closely arranged with an obscure boundary,whereas in the treated groups,a massive or focal necrosis at different levels was observed in the center and margin of tumor tissue.The relative expression levels of suvivin,VEGF and Fas protein were 3.600±0.966,4.600±0.966,4.300±0.949 respectively,in high-dose acitretin group,2.100±0.568,2.400±0.516,5.900±0.730 respectively,in combination group,5.900±1.370,6.100 ±1.1 97,2.1 00±0.568,respectively,in the negative control group,and a

  12. Mechanism of low-level ionizing radiation in inhibiting B16 melanoma blood-borne pulmonary metastasis

    International Nuclear Information System (INIS)

    Objective: To study the mechanism of low-level ionizing radiation in inhibiting B16 melanoma blood-borne pulmonary metastasis. Method: 125I-labelled B16 melanoma cells were used to investigate the effect of low dose X-irradiation on the distribution and clearance of tumor cells in mice. Results: After whole body 7.5 cGy x-irradiation, the clearance of tumor cells was significantly increased from lungs of mice. The residual numbers of tumor cells were markedly lower than those of control in blood, spleen, liver and lungs (P<0.05-0.01). The cytotoxicity of murine splenic NK cells, LTR and the phagocytosis of macrophages augmented significantly after the mice were irradiated with low dose X-rays. Conclusion: These results suggest that the enhancement of NK cytotoxicity and macrophage phagocytotic function might be one of the important reasons why low dose radiation can accelerate the clearance of tumor cells from the lungs

  13. Suppressive Effect of Juzentaihoto on Vascularization Induced by B16 Melanoma Cells In Vitro and In Vivo

    Directory of Open Access Journals (Sweden)

    Shintaro Ishikawa

    2012-01-01

    Full Text Available Juzentaihoto (JTT is well known to be one of Japanese herbal medicines, and used for the supplemental therapy of cancer patients with remarkable success. The present study, therefore, was undertaken to examine the possible therapeutic mechanisms of JTT on cancer using B16 melanoma cell (B16 cell/experimental mouse system. JTT was well mixed with rodent chow at 3.0% concentrations, and was administered orally ad libitum. Administration of JTT was started one week before tumor cell injection and continued throughout the experiment. Administration of JTT into mice significantly inhibited tumor metastasis in lungs after intravenous injection of 2×105 B16 cells in a volume of 50 μL. JTT also significantly suppressed enlargement of tumor size in hind footpad after the subcutaneous injection of 2×105 (50 μL B16 cells. In the second part of experiments, the chamber that containing B16 cells was buried in the murine back. In JTT administrated group, vascular endothelial growth factor (VEGF of chamber internal fluid significantly decreased, and vascularization of chamber circumference was also inhibited. These results strongly suggest that oral administration of JTT caused decrease in the generation of VEGF, which is responsible for vascularization, and results in inhibition of B16 cell metastasis.

  14. Immune-mediated regression of established B16F10 melanoma by intratumoral injection of attenuated Toxoplasma gondii protects against rechallenge.

    Science.gov (United States)

    Baird, Jason R; Byrne, Katelyn T; Lizotte, Patrick H; Toraya-Brown, Seiko; Scarlett, Uciane K; Alexander, Matthew P; Sheen, Mee Rie; Fox, Barbara A; Bzik, David J; Bosenberg, Marcus; Mullins, David W; Turk, Mary Jo; Fiering, Steven

    2013-01-01

    Immune recognition of tumors can limit cancer development, but antitumor immune responses are often blocked by tumor-mediated immunosuppression. Because microbes or microbial constituents are powerful adjuvants to stimulate immune responses, we evaluated whether intratumoral administration of a highly immunogenic but attenuated parasite could induce rejection of an established poorly immunogenic tumor. We treated intradermal B16F10 murine melanoma by intratumoral injection of an attenuated strain of Toxoplasma gondii (cps) that cannot replicate in vivo and therefore is not infective. The cps treatment stimulated a strong CD8(+) T cell-mediated antitumor immune response in vivo that regressed established primary melanoma. The cps monotherapy rapidly modified the tumor microenvironment, halting tumor growth, and subsequently, as tumor-reactive T cells expanded, the tumors disappeared and rarely returned. The treatment required live cps that could invade cells and also required CD8(+) T cells and NK cells, but did not require CD4(+) T cells. Furthermore, we demonstrate that IL-12, IFN-γ, and the CXCR3-stimulating cytokines are required for full treatment efficacy. The treatment developed systemic antitumor immune activity as well as antitumor immune memory and therefore might have an impact against human metastatic disease. The approach is not specific for either B16F10 or melanoma. Direct intratumoral injection of cps has efficacy against an inducible genetic melanoma model and transplantable lung and ovarian tumors, demonstrating potential for broad clinical use. The combination of efficacy, systemic antitumor immune response, and complete attenuation with no observed host toxicity demonstrates the potential value of this novel cancer therapy. PMID:23225891

  15. Melanin biosynthesis inhibitory activity of a compound isolated from young green barley (Hordeum vulgare L.) in B16 melanoma cells.

    Science.gov (United States)

    Meng, Tian Xiao; Irino, Nobuto; Kondo, Ryuichiro

    2015-07-01

    In the course to find compounds that inhibit melanin biosynthesis (i.e., whitening agents), we evaluated the effects of the methanol-soluble fraction (i.e., the water-soluble portion of methanol extracts-CHP20P-MeOH eluted fraction) from young green barley leaves on melanin production in B16 melanoma cells. Activity-guided fractionation led to an isolate called tricin (compound 1) as an inhibitory compound of melanin production in B16 melanoma cells. Furthermore, tricin analogs such as tricetin, tricetin trimethyl ether, luteolin, and apigenin were used for analyzing the structure-activity relationships (SAR) of 5,7-dihydroxyflavones studies. Tricin demonstrated stronger inhibitory activity compared to three other compounds. The results suggest that a hydroxyl group at the C-4' position and methoxy groups at the C-3',5' positions of the tricin skeleton may have important roles in this inhibitory activity in B16 melanoma cells. Our results suggest that tricin inhibits melanin biosynthesis with higher efficacy than arbutin, and it could be used as a whitening agent. PMID:25827948

  16. Inhibition of cell proliferation, migration and invasion of B16-F10 melanoma cells by α-mangostin

    Energy Technology Data Exchange (ETDEWEB)

    Beninati, Simone, E-mail: beninati@bio.uniroma2.it [Department of Biology, University “Tor Vergata”, Rome (Italy); Oliverio, Serafina [Department of Biology, University “Tor Vergata”, Rome (Italy); Cordella, Martina [Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanità, Rome (Italy); Rossi, Stefania; Senatore, Cinzia [Regina Elena National Cancer Institute, Rome (Italy); Liguori, Immacolata; Lentini, Alessandro; Piredda, Lucia [Department of Biology, University “Tor Vergata”, Rome (Italy); Tabolacci, Claudio [Department of Biology, University “Tor Vergata”, Rome (Italy); Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanità, Rome (Italy)

    2014-08-08

    Highlights: • We studied the anticancer potential of a new emerging molecule, α-mangostin (α-M). • We provide first evidences on the effects of α-M on transglutaminase activity. • We deeply examined the antimetastatic effects of α-M through many in vitro assays. • Proteomic analysis revealed that α-M promotes a reorganization at cellular level. - Abstract: In this study, we have evaluated the potential antineoplastic effects of α-mangostin (α-M), the most representative xanthone in Garcinia mangostana pericarp, on melanoma cell lines. This xanthone markedly inhibits the proliferation of high-metastatic B16-F10 melanoma cells. Furthermore, by deeply analyzing which steps in the metastatic process are influenced by xanthone it was observed that α-M strongly interferes with homotypic aggregation, adhesion, plasticity and invasion ability of B16-F10 cells, probably by the observed reduction of metalloproteinase-9 activity. The antiproliferative and antimetastatic properties of α-M have been established in human SK-MEL-28 and A375 melanoma cells. In order to identify pathways potentially involved in the antineoplastic properties of α-M, a comparative mass spectrometry proteomic approach was employed. These findings may improve our understanding of the molecular mechanisms underlying the anti-cancer effects of α-M on melanoma.

  17. Inhibition of cell proliferation, migration and invasion of B16-F10 melanoma cells by α-mangostin

    International Nuclear Information System (INIS)

    Highlights: • We studied the anticancer potential of a new emerging molecule, α-mangostin (α-M). • We provide first evidences on the effects of α-M on transglutaminase activity. • We deeply examined the antimetastatic effects of α-M through many in vitro assays. • Proteomic analysis revealed that α-M promotes a reorganization at cellular level. - Abstract: In this study, we have evaluated the potential antineoplastic effects of α-mangostin (α-M), the most representative xanthone in Garcinia mangostana pericarp, on melanoma cell lines. This xanthone markedly inhibits the proliferation of high-metastatic B16-F10 melanoma cells. Furthermore, by deeply analyzing which steps in the metastatic process are influenced by xanthone it was observed that α-M strongly interferes with homotypic aggregation, adhesion, plasticity and invasion ability of B16-F10 cells, probably by the observed reduction of metalloproteinase-9 activity. The antiproliferative and antimetastatic properties of α-M have been established in human SK-MEL-28 and A375 melanoma cells. In order to identify pathways potentially involved in the antineoplastic properties of α-M, a comparative mass spectrometry proteomic approach was employed. These findings may improve our understanding of the molecular mechanisms underlying the anti-cancer effects of α-M on melanoma

  18. Synthesis, in vitro binding and biodistribution in B16 melanoma-bearing mice of new iodine-125 spermidine benzamide derivatives

    Energy Technology Data Exchange (ETDEWEB)

    Moreau, Marie-France [INSERM UMR 484, BP 184, 63000 Clermont-Ferrand cedex (France); Univ d' Auvergne, F-63000 Clermont-Ferrand (France); Centre Jean Perrin, F-63000 Clermont-Ferrand (France); Papon, Janine [INSERM UMR 484, BP 184, 63000 Clermont-Ferrand cedex (France); Univ d' Auvergne, F-63000 Clermont-Ferrand (France); Centre Jean Perrin, F-63000 Clermont-Ferrand (France); Labarre, Pierre [INSERM UMR 484, BP 184, 63000 Clermont-Ferrand cedex (France); Univ d' Auvergne, F-63000 Clermont-Ferrand (France); Centre Jean Perrin, F-63000 Clermont-Ferrand (France); Moins, Nicole [INSERM UMR 484, BP 184, 63000 Clermont-Ferrand cedex (France); Univ d' Auvergne, F-63000 Clermont-Ferrand (France); Centre Jean Perrin, F-63000 Clermont-Ferrand (France)]. E-mail: moins@inserm484.u-clermont1.fr; Borel, Michele [INSERM UMR 484, BP 184, 63000 Clermont-Ferrand cedex (France); Univ d' Auvergne, F-63000 Clermont-Ferrand (France); Centre Jean Perrin, F-63000 Clermont-Ferrand (France); Bayle, Martine [INSERM UMR 484, BP 184, 63000 Clermont-Ferrand cedex (France); Univ d' Auvergne, F-63000 Clermont-Ferrand (France); Centre Jean Perrin, F-63000 Clermont-Ferrand (France); Bouchon, Bernadette [INSERM UMR 484, BP 184, 63000 Clermont-Ferrand cedex (France); Univ d' Auvergne, F-63000 Clermont-Ferrand (France); Centre Jean Perrin, F-63000 Clermont-Ferrand (France); Madelmont, Jean-Claude [INSERM UMR 484, BP 184, 63000 Clermont-Ferrand cedex (France); Univ d' Auvergne, F-63000 Clermont-Ferrand (France); Centre Jean Perrin, F-63000 Clermont-Ferrand (France)

    2005-05-01

    In the course of our investigations aimed at improving the biological characteristics of iodobenzamides for melanoma therapeutic applications, four new derivatives containing a spermidine chain have been prepared and radiolabeled with {sup 125}I. In vitro studies showed that all compounds displayed high affinity for melanin superior to the reference compound BZA, thus validating our experimental approach. In vivo biodistribution was investigated in B16 melanoma-bearing mice. All four compounds, particularly benzamide 3, showed accumulation in the tumor, but lower, however, than that of BZA. Moreover, high concentrations of radioactivity in other organs, namely, the liver and lung, demonstrated nonspecific tumoral uptake. In view of these results, compounds 1 2 3 4 do not appear to be suitable radiopharmaceuticals for melanoma radionuclide therapy.

  19. Evaluation of antioxidant and anti-melanogenic activities of different extracts from aerial parts of Nepeta binaludensis Jamzad in murine melanoma B16F10 cells

    Directory of Open Access Journals (Sweden)

    Zahra Tayarani-Najaran

    2016-06-01

    Conclusion: Taken together the data indicate that N. binaludensis has inhibitory activity on melanin synthesis with no cytotoxic effects in B16 melanoma cells. Therefore, it merits future investigations to apply as whitening agent in hyperpigmentation.

  20. Antiproliferative and proapoptotic actions of okra pectin on B16F10 melanoma cells

    NARCIS (Netherlands)

    Vayssade, M.; Sengkhamparn, N.; Verhoef, R.P.; Delaigue, C.; Goundiam, O.; Vigneron, P.; Voragen, A.G.J.; Schols, H.A.; Nagel, M.D.

    2010-01-01

    The proliferation and apoptosis of metastatic melanoma cells are often abnormal. We have evaluated the action of a pectic rhamnogalacturonan obtained by hot buffer extraction of okra pods (okra RG-I) on melanoma cell growth and survival in vitro. We added okra RG-I containing an almost pure RG-I car

  1. LPA is a chemorepellent for B16 melanoma cells: action through the cAMP-elevating LPA5 receptor.

    Directory of Open Access Journals (Sweden)

    Maikel Jongsma

    Full Text Available Lysophosphatidic acid (LPA, a lipid mediator enriched in serum, stimulates cell migration, proliferation and other functions in many cell types. LPA acts on six known G protein-coupled receptors, termed LPA(1-6, showing both overlapping and distinct signaling properties. Here we show that, unexpectedly, LPA and serum almost completely inhibit the transwell migration of B16 melanoma cells, with alkyl-LPA(18:1 being 10-fold more potent than acyl-LPA(18:1. The anti-migratory response to LPA is highly polarized and dependent on protein kinase A (PKA but not Rho kinase activity; it is associated with a rapid increase in intracellular cAMP levels and PIP3 depletion from the plasma membrane. B16 cells express LPA(2, LPA(5 and LPA(6 receptors. We show that LPA-induced chemorepulsion is mediated specifically by the alkyl-LPA-preferring LPA(5 receptor (GPR92, which raises intracellular cAMP via a noncanonical pathway. Our results define LPA(5 as an anti-migratory receptor and they implicate the cAMP-PKA pathway, along with reduced PIP3 signaling, as an effector of chemorepulsion in B16 melanoma cells.

  2. Phenothiazinium dyes in association with diode red laser against B16F10 melanoma cells: in vitro study

    Science.gov (United States)

    Miranda, Anderson F.; Santos, Gustavo M. P.; de Oliveira, Susana C. P. S.; Monteiro, Juliana S. C.; Sampaio, Fernando J. P.; Gomes Júnior, Rafael Araújo; Brugnera, Aldo; Gesteira, Maria F. M.; Zanin, Fátima A. A.; Pinheiro, Antônio Luiz B.; Vannier-Santos, Marcos A.

    2014-02-01

    In Brazil solar incidence is high and continuous throughout the year. Body exposure to sunlight may be a key point in the rates of individuals affected by melanoma and other types of skin cancer in many countries. Brazil already occupies the 15th place in the ranking of melanoma cases and the limitations presented by drugs used in the therapy of this cancer, new approaches are being used in an attempt to decrease the mortality of this malignancy. The aim of this study was to evaluate the effects of phenothiazinium dyes (PD) associated with laser light on murine melanoma (B16F10) in vitro by measuring cell growth using colorimetric assay before and after photodynamic therapy. We used a diode laser (λ660nm, 2.4 J/cm2, 40 mW, 60 s, CW) associated with PD at 12.5 μg/mL, time pre-irradiation of 30 minutes). The following groups were tested: control (LF-), PD (L-F+), Laser (L+F-), Laser + PD (L+F+). The results showed a significant reduction in cell growth in the group treated by the photodynamic therapy compared to the control at 24 and 48 h (p < 0.001). Were showing at 30 min PD has a dose-dependent response on B16F10 cells, but at 24 h did not demonstrated this response.

  3. Reduced paxillin expression contributes to the antimetastatic effect of 4-hydroxycoumarin on B16-F10 melanoma cells

    Directory of Open Access Journals (Sweden)

    Mandoki Juan J

    2008-05-01

    Full Text Available Abstract Background 4-Hydroxycoumarin (4-HC is a coumarin that lacks anticoagulant activity. 4-HC affects the cytoskeletal stability and decreases cell adhesion and motility of the melanoma cell line B16-F10. Together with integrins and other cytoskeletal proteins, paxillin participates in the regulation of cell adhesion and motility, acting as an adapter protein at focal adhesions. The present study determined the participation of paxillin in the reported effects of 4-HC and analyzed the role of paxillin in the formation of melanoma metastases. Results 4-HC decreased protein and mRNA levels of α- and β-paxillin isoforms in B16-F10 cells. Paxillin downregulation correlated with an inadequate translocation of paxillin to focal adhesions and a reduced phosphotyr118-paxillin pool. Consequently, 4-HC altered paxillin-mediated signaling, decreasing the phosphorylation of FAK and the level of GTP-bound Rac-1. These results partially explain the mechanism of the previously reported effects of 4-HC. Additionally, we studied the effect of 4-HC on metastatic potential of B16-F10 cells through experimental metastasis assays. In vitro treatment of cells with 4-HC inhibited their capability to originate pulmonary metastases. 4-HC did not affect cell proliferation or survival, demonstrating that its antimetastatic effect is unrelated to changes on cell viability. We also studied the importance of paxillin in metastasis by transfecting melanoma cells with paxillin-siRNA. Transfection produced a modest reduction on metastatic potential, indicating that: i paxillin plays a role as inducer of melanoma metastasis; and ii paxillin downregulation is not sufficient to explain the antimetastatic effect of 4-HC. Therefore, we evaluated other changes in gene expression by differential display RT-PCR analysis. Treatment with 4-HC produced a downregulation of Adhesion Regulating Molecule-1 (ARM-1, which correlated with a decreased adhesion of melanoma cells to lung

  4. Anti-cancer Effects of CME-1, a Novel Polysaccharide, Purified from the Mycelia of Cordyceps sinensis against B16-F10 Melanoma Cells

    Directory of Open Access Journals (Sweden)

    Thanasekaran Jayakumar

    2014-01-01

    Conclusions: These results indicate that CME-1 inhibited MMP-1 expressions in B16F10 melanoma cells through either NF-kB or ERK/p38 MAPK down regulation thereby inhibiting B16F10 cell migration. Therefore, we proposed that CME-1 might be developed as a therapeutic potential candidate for the treatment of cancer metastasis.

  5. Adoptive Transfer of Tumor-Specific Tc17 Effector T Cells Controls the Growth of B16 Melanoma in Mice

    OpenAIRE

    de la Luz Garcia-Hernandez, Maria; Hamada, Hiromasa; Reome, Joyce B.; Misra, Sara K.; Tighe, Michael P.; Dutton, Richard W.

    2010-01-01

    In vitro generated OVA-specific IL-17–producing CD8 T effector cells (Tc17) from OT-1 mice, adoptively transferred into B16-OVA tumor-bearing mice, controlled tumor growth in early and late stage melanoma. IL-17, TNF, and IFN-γ from the Tc17 effectors all played a role in an enhanced recruitment of T cells, neutrophils, and macrophages to the tumor. In addition, Tc17 cells and recently recruited, activated neutrophils produced further chemokines, including CCL3, CCL4, CCL5, CXCL9, and CXCL10,...

  6. Suppressive Effect of Juzen-Taiho-To on Lung Metastasis of B16 Melanoma Cells In Vivo

    Directory of Open Access Journals (Sweden)

    Takako Matsuda

    2011-01-01

    Full Text Available Juzen-Taiho-To (JTT is well known to be one of Kampo (Japanese herbal medicine consisted of 10 component herbs and used for the supplemental therapy of cancer patients with remarkably success. However, the precise mechanisms by which JTT could favorably modify the clinical conditions of cancer patients are not well defined. The present study, therefore, was undertaken to examine the possible mechanisms of JTT on prevention of cancer metastasis using experimental mouse model. JTT was well mixed with rodent chow at concentrations of either 0.2 or 1.0%, and administered orally ad libitum, which was started 1 week before tumor cell injection and continue throughout the experiment. Oral administration of JTT at concentration 0.2 and 1.0% into C57BL/6 male mice significantly inhibited tumor metastasis in lungs, which was induced by the intravenous injection of 2 × 105 B16 melanoma cell. JTT at a concentration of 1.0% also significantly suppressed lung metastasis of B16 melanoma cell from hind footpad in C57BL/6 mice. In the second part of experiments, the influence of the depression of natural killer (NK cell, natural killer T (NKT cell and several types of cytokines on JTT-mediated inhibition of tumor cell metastasis. Intraperitoneal injection of anti asialo-GM1 antibody against NK cells and anti NK-1.1 monoclonal antibody (mAb to NKT cells abrogated the inhibitory action of JTT on lung metastasis of B16 melanoma cells. Although intraperitoneal administration of anti-IFN-γ mAb scarcely affected the inhibitory action of JTT on tumor cell metastasis, injection of amrinone, which used for IL-12 suppression, significantly decreased the ability of JTT to prevent tumor cell metastasis. These results strongly suggest that oral administration of JTT caused increase in the production of IL-12, which is responsible for the activation of both NK cell and NKT cell, in the lungs and results in inhibition of B16 melanoma cell metastasis in the lungs.

  7. Quantification of B16 Melanoma Cells in Lungs Using Triplex Q-PCR - A New Approach to Evaluate Melanoma Cell Metastasis and Tumor Control

    DEFF Research Database (Denmark)

    Sorensen, Maria R; Pedersen, Sara R; Lindkvist, Annika;

    2014-01-01

    the outgrowth of subcutaneous melanomas. Results obtained using Q-PCR were compared to conventional counting of metastatic foci under a dissection microscope. A marked reduction in gene expression was observed in the lungs after vaccination with both vectors; however, Ad-Ii-GP showed the highest protection...... of survival once the tumor has metastasized. In the present study, we have developed a new assay for quantitative analysis of B16 melanoma metastasis in the lungs. We have used a triplex Q-PCR to determine the expression of the melanoma genes GP100/Pmel and tyrosinase-related protein 2 (TRP-2), and found......, and matching results were obtained by enumeration of visible tumor nodules on the lung surfaces. Finally, we could show that inhibition of tumor metastasis required antigen-specific CD8 T cells and IFNγ, but not perforin. In conclusion, the presented results validate triplex Q-PCR as a fast, objective...

  8. In vitro evaluation of low-intensity light radiation on murine melanoma (B16F10) cells.

    Science.gov (United States)

    Peidaee, P; Almansour, N M; Pirogova, E

    2016-03-01

    Changes in the energy state of biomolecules induced by electromagnetic radiation lead to changes in biological functions of irradiated biomolecules. Using the RRM approach, it was computationally predicted that far-infrared light irradiation in the range of 3500-6000 nm affects biological activity of proto-oncogene proteins. This in vitro study evaluates quantitatively and qualitatively the effects of selected far-infrared exposures in the computationally determined wavelengths on mouse melanoma B16F10 cells and Chinese hamster ovarian (CHO) cells by MTT (thiazolyl blue tetrazolium bromide) cell proliferation assay and confocal laser-scanning microscopy (CLSM). This paper also presents the findings obtained from irradiating B16F10 and CHO cells by the selected wavelengths in visible and near-infrared range. The MTT results show that far-infrared wavelength irradiation induces detrimental effect on cellular viability of B16F10 cells, while that of normal CHO cells is not affected considerably. Moreover, CLSM images demonstrate visible cellular detachment of cancer cells. The observed effects support the hypothesis that far-infrared light irradiation within the computationally determined wavelength range induces biological effect on cancer cells. From irradiation of selected visible and near-infrared wavelengths, no visible changes were detected in cellular viability of either normal or cancer cells.

  9. In vitro evaluation of low-intensity light radiation on murine melanoma (B16F10) cells.

    Science.gov (United States)

    Peidaee, P; Almansour, N M; Pirogova, E

    2016-03-01

    Changes in the energy state of biomolecules induced by electromagnetic radiation lead to changes in biological functions of irradiated biomolecules. Using the RRM approach, it was computationally predicted that far-infrared light irradiation in the range of 3500-6000 nm affects biological activity of proto-oncogene proteins. This in vitro study evaluates quantitatively and qualitatively the effects of selected far-infrared exposures in the computationally determined wavelengths on mouse melanoma B16F10 cells and Chinese hamster ovarian (CHO) cells by MTT (thiazolyl blue tetrazolium bromide) cell proliferation assay and confocal laser-scanning microscopy (CLSM). This paper also presents the findings obtained from irradiating B16F10 and CHO cells by the selected wavelengths in visible and near-infrared range. The MTT results show that far-infrared wavelength irradiation induces detrimental effect on cellular viability of B16F10 cells, while that of normal CHO cells is not affected considerably. Moreover, CLSM images demonstrate visible cellular detachment of cancer cells. The observed effects support the hypothesis that far-infrared light irradiation within the computationally determined wavelength range induces biological effect on cancer cells. From irradiation of selected visible and near-infrared wavelengths, no visible changes were detected in cellular viability of either normal or cancer cells. PMID:26002595

  10. Inhibitory effects of Morinda citrifolia extract and its constituents on melanogenesis in murine B16 melanoma cells.

    Science.gov (United States)

    Masuda, Megumi; Itoh, Kimihisa; Murata, Kazuya; Naruto, Shunsuke; Uwaya, Akemi; Isami, Fumiyuki; Matsuda, Hideaki

    2012-01-01

    The objective of this study was to examine the effects of Morinda citrifolia (noni) extract and its constituents on α-melanocyte stimulating hormone (α-MSH)-stimulated melanogenesis in cultured murine B16 melanoma cells (B16 cells). A 50% ethanolic extract of noni seeds (MCS-ext) showed significant inhibition of melanogenesis with no effect on cell proliferation. MCS-ext was more active than noni leaf and fruit flesh extracts. Activity guided fractionation of MCS-ext led to the isolation of two lignans, 3,3'-bisdemethylpinoresinol (1) and americanin A (2), as active constituents. To elucidate the mechanism of melanogenesis inhibition by the lignans, α-MSH-stimulated B16 cells were treated with 1 (5 μM) and 2 (200 μM). Time-dependent increases of intracellular melanin content and tyrosinase activity, during 24 to 72 h, were inhibited significantly by treatment with the lignans. The activity of 1 was greater than that of 2. Western blot analysis suggested that the lignans inhibited melanogenesis by down regulation of the levels of phosphorylation of p38 mitogen-activated protein kinase, resulting in suppression of tyrosinase expression.

  11. Inhibitory effects of whisky polyphenols on melanogenesis in mouse B16 melanoma cells.

    Science.gov (United States)

    Yoshioka, Sayaka; Terashita, Takao; Yoshizumi, Hajime; Shirasaka, Norifumi

    2011-01-01

    Whisky exerts an inhibitory effect on melanogenesis in B16 cells, the anti-melanogenic activity being positively correlated with the aging period and anti-oxidative activity of whisky. We examined the correlation between the inhibition of melanogenesis and the concentration of each compound in various whiskies to evaluate the importance of 11 different whisky polyphenols, including ellagic acid, gallic acid and lyoniresinol, in the anti-melanogenic activity of whisky. The concentration of all the compounds was positively correlated with the anti-melanogenic activity of whisky. Ellagic acid, gallic acid and lyoniresinol were the predominant polyphenols in the whiskies measured by HPLC. These three compounds also significantly inhibited the melanogenesis and tyrosinase activity in B16 cells. Ellagic acid, gallic acid and lyoniresinol were confirmed as the major participants in the anti-melanogenic activity of whisky.

  12. Oligoesculin fraction induces anti-tumor effects and promotes immune responses on B16-F10 mice melanoma.

    Science.gov (United States)

    Mokdad Bzeouich, Imen; Mustapha, Nadia; Sassi, Aicha; Ghedira, Kamel; Ghoul, Mohamed; Chebil, Latifa; Luis, José; Chekir-Ghedira, Leila

    2016-08-01

    Laccase was used to enzymatically polymerize esculin. Oligoesculin fraction was obtained after ultrafiltration through a 5-kDa membrane. Several studies have been carried out to prove the effectiveness of natural substances such as immunomodulators to promote the anti-cancer activity in situ. The purpose of our report was to explore whether the anti-tumor potential of the oligoesculin fraction in vitro and in vivo is linked to its immunological mechanisms in melanoma-bearing mice. We revealed that oligoesculin fraction reduced B16-F10 proliferation and migration in vitro in a dose-related manner. Moreover, melanin synthesis and tyrosinase activity were inhibited in these melanoma cells in a concentration-dependent way. The anti-tumor potential of oligoesculin fraction was also assessed in vivo. Our results showed that intraperitoneal administration of oligoesculin fraction, at 50 mg/kg body weight (b.w.) for 21 days, reduced tumor size and weight with percentages of inhibition of 94 and 87 %, respectively. Oligoesculin fraction was effective in promoting lysosomal activity and nitric oxide (NO) production by peritoneal macrophages in tumor-implanted mice. In addition, the activities of natural killer (NK), cytotoxic T lymphocytes, and macrophages were significantly enhanced by oligoesculin fraction. These findings suggested that this polymer with its anti-tumor and immunomodulatory properties could be used for the treatment of melanoma. PMID:26960691

  13. Diarylheptanoids with inhibitory effects on melanogenesis from the rhizomes of Curcuma comosa in B16 melanoma cells.

    Science.gov (United States)

    Matsumoto, Takahiro; Nakamura, Seikou; Nakashima, Souichi; Yoshikawa, Masayuki; Fujimoto, Katsuyoshi; Ohta, Tomoe; Morita, Azumi; Yasui, Rie; Kashiwazaki, Eri; Matsuda, Hisashi

    2013-09-15

    The methanolic extract from the dried rhizomes of Curcuma comosa cultivated in Thailand was found to inhibit melanogenesis in theophylline-stimulated murine B16 melanoma 4A5 cells. From the methanolic extract, three new diarylheptanoids, diarylcomosols I-III, were isolated together with 12 known diarylheptanoids. Their chemical structures were elucidated on the basis of chemical and physicochemical evidence. The diarylheptanoids inhibited melanogenesis, and several structural requirements of the active constituents for the inhibition were clarified. In particular, (3R)-1,7-bis(4-hydroxyphenyl)-(6E)-6-hepten-3-ol exhibited stronger inhibitory effect [IC50=0.36 μM] without inducing cytotoxicity. The biological effect was much stronger than that of a reference compound, arbutin [IC50=174 μM]. We conclude that diarylheptanoid analogs are promising therapeutic agents for the treatment of skin disorders.

  14. Inhibitors of melanogenesis in B16 melanoma 4A5 cells from flower buds of Lawsonia inermis (Henna).

    Science.gov (United States)

    Nakashima, Souichi; Oda, Yoshimi; Nakamura, Seikou; Liu, Jiang; Onishi, Koko; Kawabata, Miki; Miki, Hisako; Himuro, Yugo; Yoshikawa, Masayuki; Matsuda, Hisashi

    2015-07-01

    The methanolic extract of Lawsonia inermis L. (henna) showed significant inhibitory activity toward melanogenesis in B16 melanoma 4A5 cells. Among the constituents isolated from the methanolic extract, luteolin, quercetin, and (±)-eriodictyol showed stronger inhibitory activity than the reference compound, arbutin. Several structure-activity relationships of the flavonoids were suggested, and OGlc

  15. Hinokitiol Inhibits Melanogenesis via AKT/mTOR Signaling in B16F10 Mouse Melanoma Cells.

    Science.gov (United States)

    Tsao, Yu-Tzu; Huang, Yu-Fen; Kuo, Chun-Yu; Lin, Yu-Chiang; Chiang, Wei-Cheng; Wang, Wei-Kuang; Hsu, Chia-Wei; Lee, Che-Hsin

    2016-01-01

    H inokitiol purified from the heartwood of cupressaceous plants has had various biological functions of cell differentiation and growth. Hinokitiol has been demonstrated as having an important role in anti-inflammation and anti-bacteria effect, suggesting that it is potentially useful in therapies for hyperpigmentation. Previously, hinokitiol inhibited the production of melanin by inhibiting tyrosinase activity. The autophagic signaling pathway can induce hypopigmentation. This study is warranted to investigate the mechanism of hinokitiol-induced hypopigmentation through autophagy in B16F10 melanoma cells. The melanin contents and expression of microthphalmia associated transcription factor (MITF) and tyrosinase were inhibited by treatment with hinokitiol. Moreover, the phosphorylation of the protein express levels of phospho-protein kinase B (P-AKT) and phospho-mammalian targets of rapamycin (P-mTOR) were reduced after hinokitiol treatment. In addition, the microtubule associated protein 1 light chain 3 (LC3) -II and beclin 1 (autophagic markers) were increased after the B16F10 cell was treated with hinokitiol. Meanwhile, hinokitiol decreased cellular melanin contents in a dose-dependent manner. These findings establish that hinokitiol inhibited melanogenesis through the AKT/mTOR signaling pathway. PMID:26901194

  16. Influence of Fuse-2 Recipe on Mice B16 Melanoma Cells Tyrosinase%复色2号方对鼠黑素瘤B16细胞酪氨酸酶的影响

    Institute of Scientific and Technical Information of China (English)

    杨柳; 张明明; 马玲玲

    2012-01-01

    Objective; To explore molecular mechanism of fuse - 2 recipe on treating vitiligo. Methods; To determine the influence of fuse -2 recipe on B16 cell proliferation by MTT method, measuring and recording B16 cell proliferation by the microplate reader. To determine the influence of fuse -2 recipe on B16 cell tyrosines activity by tyrosinase DOPA rate oxidation method. To determine the melanin content of B16 cell by NaOH decomposition method. Results: There was no statistical significance(P >0.05)of fuse -2 drug concentration at 1 ~ lOOOg/L on B16 cell proliferation,but there was statistical significance (P <0. 05) on tyrosinase activity and melanin synthesis in a concentration dependent manner. Conclusion :Without effect on B16 melanoma cell proliferation, fuse -2 has a strong correlation effect during the dose on tyrosinase activity and melanin synthesis, which could achieve the goal of the treating vitiligo.%目的:探讨复色2号方治疗白癜风的分子学作用机制.方法:采用MTT法测定复色2号方对B16细胞增值的影响,在酶标仪下测定并记录B16细胞增殖情况;采用酪氨酸酶多巴速率氧化法测定复色2号方对酪氨酸酶活性的影响;采用NaOH裂解法测定B16细胞黑素含量.结果:复色2号方药物浓度在1~1000g/L对黑素瘤B16细胞增殖无统计学意义(P>0.05),但对酪氨酸酶活性和黑素合成有统计学意义(P<0.05),呈浓度依赖性.结论:复色2号方对黑素瘤B16细胞增殖无影响,但对酪氨酸酶活性和黑素合成有较强的剂量相关性促进作用,可以达到治疗白癜风的目的.

  17. Sarcophine-Diol, a Skin Cancer Chemopreventive Agent, Inhibits Proliferation and Stimulates Apoptosis in Mouse Melanoma B16F10 Cell Line

    OpenAIRE

    Hesham Fahmy; Ahmed, Safwat A.; Szymanski, Pawel T.; Bhimanna Kuppast; Sherief Khalifa

    2011-01-01

    Sarcodiol (SD) is a semi-synthetic derivative of sarcophine, a marine natural product. In our previous work, we reported the significant chemopreventive effects of SD against non-melanoma skin cancer both in vitro and in vivo mouse models. In this investigation, we extended this work to study the effect of sarcodiol on melanoma development, the more deadly form of skin cancer, using the mouse melanoma B16F10 cell line. In this study we report that SD inhibits the de novo DNA synthesis and enh...

  18. Improving anticancer efficacy of (--epigallocatechin-3-gallate gold nanoparticles in murine B16F10 melanoma cells

    Directory of Open Access Journals (Sweden)

    Chen CC

    2014-05-01

    Full Text Available Cheng-Cheung Chen,1,2 Dar-Shih Hsieh,1,3 Kao-Jean Huang,4 Yi-Lin Chan,5 Po-Da Hong,6 Ming-Kung Yeh,6–8,* Chang-Jer Wu1,*1Department of Food Science, National Taiwan Ocean University, Keelung, 2Institute of Preventive Medicine, National Defense Medical Center, Taipei, 3Division of Urology, Department of Surgery, Ren-Ai Hospital, Shulin, New Taipei City, 4Department of Life Science and Institute of Biotechnology, National Dong Hwa University, Hualien, 5Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, 6Materials Technology Program, Graduate Institute of Applied Science and Technology, National Taiwan University of Science and Technology, Taipei, 7School of Pharmacy, National Defense Medical Center, Taipei, 8Food and Drug Administration, Ministry of Health and Welfare, Taipei, Taiwan, Republic of China*These authors contributed equally to this workAbstract: (--Epigallocatechin-3-gallate (EGCG, the major bioactive constituent in green tea, has been reported to effectively inhibit the formation and development of tumors. To maximize the effectiveness of EGCG, we attached it to nanogold particles (EGCG-pNG in various ratios to examine in vitro cytotoxicity and in vivo anti-cancer activity. EGCG-pNG showed improved anti-cancer efficacy in B16F10 murine melanoma cells; the cytotoxic effect in the melanoma cells treated with EGCG-pNG was 4.91 times higher than those treated with EGCG. The enhancement is achieved through mitochondrial pathway-mediated apoptosis as determined by annexin V assay, JC-10 staining, and caspase-3, -8, -9 activity assay. Moreover, EGCG-pNG was 1.66 times more potent than EGCG for inhibition of tumor growth in a murine melanoma model. In the hemolysis assay, the pNG surface conjugated with EGCG is most likely the key factor that contributes to the decreased release of hemoglobin from human red blood cells.Keywords: gold nanoparticles, EGCG, anticancer, melanoma

  19. Effector cells derived from naive T cells used in tumor immunotherapy of mice bearing B16 melanoma

    Institute of Scientific and Technical Information of China (English)

    Wen Ming; Xu Weili; Ren Lili; Gao Fei; Cui Naipeng; Wen Junye; Li Xinjiang

    2014-01-01

    Background Adoptive cell transfer (ACT) immunotherapy has been used clinically for years to treat malignancies.Improving the killing efficiency of effector cells,such as tumor-specific cytotoxic T lymphocytes (CTLs),is an important component for enhancing the clinical response of cancer immunotherapy.Hence,we explored a novel method for preparing cancer-specific CTLs using naive T lymphocytes.Methods C57BL/6 mice bearing B16 melanoma tumors were pretreated with cyclophosphamide (CTX) by peritoneal injection.The immunosuppressive influence of CTX on tumor regression and the tumor microenvironment was assessed.Naive T cells and T cell pools were isolated via negative selection using immunomagnetic beads.The proliferative potential and cytokine production of different T cell subpopulations were evaluated in vitro.Tumor-specific CTLs derived from naive T cells (naive CD4+ T cells:naive CD8+ T cells=2:1) and pooled T cells were generated in vitro,respectively.B16 melanoma-bearing C57BL/6 mice were pretreated with CTX,followed by ACT immunotherapy using dendritic cell-induced CTLs.The homing abilities of the effector cells and interleukin-2 (IL-2),interferon-y,granzyme B,and perforin mRNA levels in tumor tissues were evaluated,and the change in tumor volume was measured.Results Mice receiving CTX peritoneal pretreatment injections did not display tumor regression compared with control mice.However,a significant downregulation of splenic Tregs and tumor growth factor-β1 (TGF-β1) and interleukin-10 (IL-10) serum levels was observed (P <0.05).Naive T cells showed a stronger proliferative capacity and elevated cytokine production than did pooled T cells (P <0.05).In addition,effector cells generated from naive T cells displayed more potent antitumor activity in vivo than those derived from pooled T cells (P <0.05).Conclusion Effector cells derived from the naive T cells possess a stronger proliferative potential,homing capacity,and enhanced cytokine production

  20. Arthrophytum scoparium inhibits melanogenesis through the down-regulation of tyrosinase and melanogenic gene expressions in B16 melanoma cells.

    Science.gov (United States)

    Chao, Hui-Chia; Najjaa, Hanen; Villareal, Myra O; Ksouri, Riadh; Han, Junkyu; Neffati, Mohamed; Isoda, Hiroko

    2013-02-01

    Melanin performs a crucial role in protecting the skin against harmful ultraviolet light. However, hyperpigmentation may lead to aesthetic problems and disorders such as solar lentigines (SL), melasma, postinflammatory hyperpigmentation and even melanoma. Arthrophytum scoparium grows in the desert in the North African region, and given this type of environment, A. scoparium exhibits adaptations for storing water and produces useful bioactive factors. In this study, the effect of A. scoparium ethanol extract (ASEE) on melanogenesis regulation in B16 murine melanoma cells was investigated. Cells treated with 0.017% (w/v) ASEE showed a significant inhibition of melanin biosynthesis in a time-dependent manner without cytotoxicity. To clarify the mechanism behind the ASEE-treated melanogenesis regulation, the expressions of tyrosinase enzyme and melanogenesis-related genes were determined. Results showed that the expression of tyrosinase enzyme was significantly decreased and Tyr, Trp-1, Mitf and Mc1R mRNA expressions were significantly down-regulated. LC-ESI-TOF-MS analysis of the extract identified the presence of six phenolic compounds: coumaric acid, cinnamic acid, chrysoeriol, cyanidin, catechol and caffeoylquinic acid. The melanogenesis inhibitory effect of ASEE may therefore be attributed to its catechol and tetrahydroisoquinoline derivative content. We report here that ASEE can inhibit melanogenesis in a time-dependent manner by decreasing the tyrosinase protein and Tyr, Trp-1, Mitf and Mc1R mRNA expressions. This is the first report on the antimelanogenesis effect of A. scoparium and on its potential as a whitening agent. PMID:23362872

  1. Neem leaf glycoprotein optimizes effector and regulatory functions within tumor microenvironment to intervene therapeutically the growth of B16 melanoma in C57BL/6 mice

    Directory of Open Access Journals (Sweden)

    Subhasis Barik

    2015-01-01

    Full Text Available Therapy with neem leaf glycoprotein (NLGP inhibits murine B16-melanoma in vivo and improves survivability. Studies on tumor-microenvironment (TME from NLGP treated mice (NLGP-TME suggests that anti-tumor effect is directly associated with enhanced CD8+T cell activity, dominance of type 1 cytokines/chemokine network with downregulation of suppressive cellular functions. NLGP-TME educated CD8+T cells showed higher perforin and granzymeB expression with greater in vitro cytotoxicity against B16 melanoma. These CD8+T cells showed proportionally lower FasR expression, denotes prevention from activation induced cell death by NLGP. Accumulated evidences strongly suggest NLGP influenced normalized TME allows CD8+T cells to perform optimally to inhibit melanoma growth.

  2. Vasoactive intestinal peptide stimulates melanogenesis in B16F10 mouse melanoma cells via CREB/MITF/tyrosinase signaling.

    Science.gov (United States)

    Yuan, Xing-Hua; Yao, Cheng; Oh, Jang-Hee; Park, Chi-Hyun; Tian, Yu-Dan; Han, Mira; Kim, Ji Eun; Chung, Jin Ho; Jin, Zhe-Hu; Lee, Dong Hun

    2016-08-26

    Vasoactive intestinal peptide (VIP), one of the major skin neuropeptides, has been suggested to have active roles in the pathogenesis of inflammatory skin disorders such as atopic dermatitis and psoriasis, which can commonly cause post-inflammatory hyperpigmentation. However, the effect of VIP on melanogenesis remains unknown. In this study, we showed that the melanin contents, tyrosinase activity, and gene expression of tyrosinase and microphthalmia-associated transcription factor (MITF) were significantly increased by treatment with VIP in B16F10 mouse melanoma cells and the stimulatory melanogenic effect was further examined in human epidermal melanocytes (HEMns). In addition, phosphorylated levels of CRE-binding protein (CREB) and protein kinase A (PKA) were markedly increased after VIP treatment, but not p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase (ERK), or Akt, indicating the possible PKA-CREB signaling pathway involved in VIP-induced melanogenesis. This result was further verified by the fact that VIP induced increased melanin synthesis, and protein levels of phosphorylated CREB, MITF, tyrosinase were significantly attenuated by H89 (a specific PKA inhibitor). These data suggest that VIP-induced upregulation of tyrosinase through the CREB-MITF signaling pathway plays an important role in finding new treatment strategy for skin inflammatory diseases related pigmentation disorders. PMID:27343558

  3. Sarcophine-Diol, a Skin Cancer Chemopreventive Agent, Inhibits Proliferation and Stimulates Apoptosis in Mouse Melanoma B16F10 Cell Line

    Directory of Open Access Journals (Sweden)

    Hesham Fahmy

    2011-12-01

    Full Text Available Sarcodiol (SD is a semi-synthetic derivative of sarcophine, a marine natural product. In our previous work, we reported the significant chemopreventive effects of SD against non-melanoma skin cancer both in vitro and in vivo mouse models. In this investigation, we extended this work to study the effect of sarcodiol on melanoma development, the more deadly form of skin cancer, using the mouse melanoma B16F10 cell line. In this study we report that SD inhibits the de novo DNA synthesis and enhances fragmentation of DNA. We also evaluated the antitumor effect of SD on melanoma cell viability using several biomarkers for cell proliferation and apoptosis. SD inhibits the expression levels of signal transducers and activators of transcription protein (STAT-3 and cyclin D1, an activator of cyclin-dependent kinase 4 (Cdk4. SD treatment also enhances cellular level of tumor suppressor protein 53 (p53 and stimulates cleavage of the nuclear poly (ADP-ribose polymerase (cleaved-PARP. SD also enhances cellular levels of cleaved Caspase-3, -8, -9 and stimulates enzymatic activities of Caspase-3, -8 and -9. These results, in addition to inhibition of cell viability, suggest that SD inhibits melanoma cell proliferation by arresting the cell-division cycle in a Go quiescent phase and activates programmed cell death (apoptosis via extrinsic and intrinsic pathways. Finally, these studies demonstrate that SD shows a very promising chemopreventive effect in melanoma B16F10 tumor cells.

  4. Pyrostegia venusta heptane extract containing saturated aliphatic hydrocarbons induces apoptosis on B16F10-Nex2 melanoma cells and displays antitumor activity in vivo

    Science.gov (United States)

    Figueiredo, Carlos R.; Matsuo, Alisson L.; Pereira, Felipe V.; Rabaça, Aline N.; Farias, Camyla F.; Girola, Nátalia; Massaoka, Mariana H.; Azevedo, Ricardo A.; Scutti, Jorge A.B.; Arruda, Denise C.; Silva, Luciana P.; Rodrigues, Elaine G.; Lago, João Henrique G.; Travassos, Luiz R.; Silva, Regildo M.G.

    2014-01-01

    Background: Pyrostegia venusta (Ker. Gawl.) Miers (Bignoniacea) is a medicinal plant from the Brazilian Cerrado used to treat leucoderma and common diseases of the respiratory system. Objective: To investigate the antitumor activity of P.venusta extracts against melanoma. Materials and Methods: The cytotoxic activity and tumor induced cell death of heptane extract (HE) from P. venusta flowers was evaluated against murine melanoma B16F10-Nex2 cells in vitro and in a syngeneic model in vivo. Results: We found that HE induced apoptosis in melanoma cells by disruption of the mitochondrial membrane potential, induction of reactive oxygen species and late apoptosis evidenced by plasma membrane blebbing, cell shrinkage, chromatin condensation and DNA fragmentation, exposure of phosphatidylserine on the cell surface and activation of caspase-2,-3,-8,-9. HE was also protective against singeneyc subcutaneous melanoma HE compounds were also able to induce cell cycle arrest at G2/M phases on tumor cells. On fractionation of HE in silica gel we isolated a cytotoxic fraction that contained a mixture of saturated hydrocarbons identified by 1H NMR and GC-MS analyses. Predominant species were octacosane (C28H58-36%) and triacontane (C30H62-13%), which individually showed significant cytotoxic activity against murine melanoma B16F10-Nex2 cells in vitro and a very promising antitumor protection against subcutaneous melanoma in vivo. Conclusion: The results suggest that the components of the heptane extract, mainly octasane and triacontane, which showed antitumor properties in experimental melanoma upon regional administration, might also be therapeutic in human cancer, such as in the mostly epidermal and slowly invasive melanomas, such as acral lentiginous melanoma, as an adjuvant treatment to surgical excision. PMID:24991116

  5. Pyrostegia venusta heptane extract containing saturated aliphatic hydrocarbons induces apoptosis on B16F10-Nex2 melanoma cells and displays antitumor activity in vivo

    Directory of Open Access Journals (Sweden)

    Carlos R Figueiredo

    2014-01-01

    Full Text Available Background: Pyrostegia venusta (Ker. Gawl. Miers (Bignoniacea is a medicinal plant from the Brazilian Cerrado used to treat leucoderma and common diseases of the respiratory system. Objective: To investigate the antitumor activity of P.venusta extracts against melanoma. Materials and Methods: The cytotoxic activity and tumor induced cell death of heptane extract (HE from P. venusta flowers was evaluated against murine melanoma B16F10-Nex2 cells in vitro and in a syngeneic model in vivo. Results: We found that HE induced apoptosis in melanoma cells by disruption of the mitochondrial membrane potential, induction of reactive oxygen species and late apoptosis evidenced by plasma membrane blebbing, cell shrinkage, chromatin condensation and DNA fragmentation, exposure of phosphatidylserine on the cell surface and activation of caspase-2,-3,-8,-9. HE was also protective against singeneyc subcutaneous melanoma HE compounds were also able to induce cell cycle arrest at G2/M phases on tumor cells. On fractionation of HE in silica gel we isolated a cytotoxic fraction that contained a mixture of saturated hydrocarbons identified by 1 H NMR and GC-MS analyses. Predominant species were octacosane (C 28 H 58 -36% and triacontane (C 30 H 62 -13%, which individually showed significant cytotoxic activity against murine melanoma B16F10-Nex2 cells in vitro and a very promising antitumor protection against subcutaneous melanoma in vivo. Conclusion: The results suggest that the components of the heptane extract, mainly octasane and triacontane, which showed antitumor properties in experimental melanoma upon regional administration, might also be therapeutic in human cancer, such as in the mostly epidermal and slowly invasive melanomas, such as acral lentiginous melanoma, as an adjuvant treatment to surgical excision.

  6. Melanoma in Organ Transplant Recipients: Incidence, Outcomes and Management Considerations

    Directory of Open Access Journals (Sweden)

    Faisal R. Ali

    2012-01-01

    Full Text Available The incidence of melanoma continues to increase year on year. With better surgical techniques and medical management, greater numbers of organ transplants are being performed annually with much longer graft survival. The authors review our current understanding of the incidence of melanoma amongst organ transplant recipients, outcomes compared to the immunocompetent population, and management strategies in this burgeoning group.

  7. Anti-tumour efficacy of mouse spleen cells separated with Dolichos biflorus lectin (DBA) in experimental pulmonary metastasis of B16 melanoma cells.

    OpenAIRE

    Okada, T.; Higuchi, M.; Takano, M; Maruyama, T.; Imai, Y; Osawa, T

    1990-01-01

    Anti-tumour effector cells were generated through 4 days culture of normal C57BL/6 splenocytes in a medium containing concanavalin A supernatant and then fractionated with Dolichos biflorus lectin (DBA) into DBA+ (agglutinable with DBA) and DBA- (non-agglutinable with DBA) cells. The DBA- cells, infused intravenously into mice together with B16 melanoma cells, or adoptively transferred into mice 3 days after the injection of B16 cells, caused a marked decrease in the number of lung nodules, w...

  8. 姜辣素对小鼠 B16细胞黑素合成的影响%Effects of Gingerol on Melanogenesis of B16 Melanoma Cells

    Institute of Scientific and Technical Information of China (English)

    神芳丽; 霍仕霞

    2014-01-01

    Objective To study the effects of the gingerol on the melanogenesis in melanoma B16 cells. Methods Melanoma cells were cultured with gingerol at 12. 5, 25. 0, 50. 0, 100. 0, 200. 0 μmol · L-1 and positive control drug hydroquinone,respectively,using Dulbecco's modified eagle's medium(DMEM) as the blank control group. The cell proliferation was measured by methyl thiazolyltet tetrazolium ( MTT) colorimetric assay. The tyrosinase activity and melanin content were measured by colorimetry assay. Results Gingerol at different concentrations had inhibitory effect on B16 cell proliferation compared with the blank control group ( P 48%。与空白对照组比较,各浓度姜辣素均能够显著降低酪氨酸酶活性(P25.0μmol·L-1时,抑制黑素含量的水平基本无变化。结论姜辣素能有效抑制黑素细胞增殖和降低酪氨酸酶活性和黑素含量,从而减少黑素合成。

  9. Antitumor Effects In Vitro and In Vivo and Mechanisms of Protection against Melanoma B16F10-Nex2 Cells By Fastuosain, a Cysteine Proteinase from Bromelia fastuosa

    OpenAIRE

    Guimarães-Ferreira, Carla A; Rodrigues, Elaine G; Renato A Mortara; Hamilton Cabral; Fabiana A. Serrano; Ricardo Ribeiro-dos-Santos; Travassos, Luiz R.

    2007-01-01

    In the present work, the antitumor effect of fastuosain, a cysteine proteinase from Bromelia fastuosa, was investigated. In the intravenous model of lung colonization in C57BI/6 mice, fastuosain and bromelain injected intraperitoneally were protective, very few nodules of B16F10-Nex2 melanoma cells were detected. Tumor cells treated with fastuosain showed reduced expression of CD44 and decreased invasion through Matrigel, lost their cytoplasmic extensions and substrate adherence, became round...

  10. BFD-22 a new potential inhibitor of BRAF inhibits the metastasis of B16F10 melanoma cells and simultaneously increased the tumor immunogenicity.

    Science.gov (United States)

    Ferreira, Adilson Kleber; Pasqualoto, Kerly Fernanda Mesquita; Kruyt, Frank A E; Palace-Berl, Fanny; Azevedo, Ricardo Alexandre; Turra, Kely Medeiros; Rodrigues, Cecilia Pessoa; Ferreira, Ana Carolina Franco; Salomón, Maria Alejandra Clavijo; de Sá Junior, Paulo Luiz; Farias, Camyla Fernandes; Figueiredo, Carlos Rogerio; Tavares, Leoberto Costa; Barbuto, José Alexandre Marzagão; Jorge, Salomão Dória

    2016-03-15

    Benzofuroxan is an interesting ring system, which has shown a wide spectrum of biological responses against tumor cell lines. We investigated, herein, the antitumor effects of benzofuroxan derivatives (BFDs) in vitro and in a melanoma mouse model. Cytotoxic effects of twenty-two BFDs were determined by MTT assay. Effects of BFD-22 in apoptosis and cell proliferation were evaluated using Annexin V-FITC/PI and CFSE staining. In addition, the effects in the cell cycle were assessed. Flow cytometry, western blot, and fluorescence microscopy analysis were employed to investigate the apoptosis-related proteins and the BRAF signaling. Cell motility was also exploited through cell invasion and migration assays. Molecular docking approach was performed in order to verify the BFD-22 binding mode into the ATP catalytic site of BRAF kinase. Moreover, the BFD-22 antitumor effects were evaluated in a melanoma murine model using B16F10. BFD-22 was identified as a potential hit against melanoma cells. BFD-22 induced apoptosis and inhibited cell proliferation of B16F10 cells. BFD-22 has suppressed, indeed, the migratory and invasive behavior of B16F10 cells. Cyclin D1 and CDK4 expression were reduced leading to cell cycle arrest at G0/G1 phase. Of note, phosphorylation of BRAF at Ser338 was strongly down-regulated by BFD-22 in B16F10 cells. The accommodation/orientation into the binding site of BRAF was similar of BAY43-9006 (co-crystallized inhibitor of BRAF, sorafenib). Importantly, BFD-22 presented in vivo antimetastatic effects and showed better therapeutic efficacy than sorafenib and taxol. BFD-22 can be considered as a new lead compound and, then, can be helpful for the designing of novel drug candidates to treat melanoma. PMID:26876618

  11. Enhancement of pEgr-p16 expression induced by irradiation and its anti-tumor effect on B16 melanoma cells in vitro

    International Nuclear Information System (INIS)

    Objective: To study the antitumour effect of irradiation combination with recombined pEgr-p16 plasmid on melanoma B16 cells. Methods: pEgr-p16 plasmids were transfected into B16 cell line. Quantitative RT-PCR, flow cytometry and MTT methods were used to detect the expression of p16, cell apoptosis and inhibition effect, respectively. Results: The p16 expressions in different doses X-ray irradiation group were about 3.78-6.67 times higher than that in sham-irradiation group (P<0.01). The expressions of p16 gradually increased with time after exposure to 2 Gy X-ray irradiation and reached to maximum at 72 h. Apoptosis rate in transfected p16 combined with irradiation group was higher than that in either plasmid or irradiation group alone (P<0.05-0.01). The number of pEgr-p16-transfected B16 cells exposed to 2 Gy X-ray irradiation was significantly less than those in other experimental groups (P<0.05-0.001). Conclusions: pEgr-p16 gene transfection combined with irradiation could suppress melanoma B16 cell growth. And the inhibition was more effective than those in gene- or irradiation-therapy alone. (authors)

  12. The Mechanism of Medium Stimulating the Melanin Synthesis in B16 Murine Melanoma Cells%培养基诱导B16细胞黑色素合成及机理研究

    Institute of Scientific and Technical Information of China (English)

    陈金妹; 林进妹; 黄家福; 欧一新; 曹娜; 张秀芬; 朱祯慧; 潘裕添

    2016-01-01

    研究不同培养基对B16细胞体外培养生物学特性的影响及其黑色素表达能力与相关分子作用机制。方法:利用不同培养基对B16细胞进行体外连续传代培养,考查培养基对细胞形态、细胞成活率、细胞周期和黑色素合成等方面的影响。同时,借助RT-PCR、Western Blot和双向电泳等技术,研究与黑色素生物合成相关蛋白质的表达水平。结果:B16细胞在Gibco-RPMI-1640-31800和Gibco-DMEM-12800这两种培养基中培养时,细胞外观形态良好、细胞活力强、传代周期短并且均可稳定连续传代培养,两者的细胞周期也一致;B16细胞在Gibco-RPMI-1640-31800培养基中基本不合成黑色素,而在Gibco-DMEM-12800培养基能大量合成黑色素。分析B16细胞中三种黑色素合成相关蛋白质(Tyrosinase,Tyrosinase-related protein 1和Tyrosinase-related protein 2)在以上两种培养基中表达的差异,发现这三种蛋白质的mRNA转录水平基本没有差异,在Gibco-DMEM-12800培养基中这三种蛋白质的翻译水平均明显提高,总蛋白质组也更加多样。结论:Gibco-DMEM-12800培养基通过提高Tyrosinase、TRP1和TRP2等三种蛋白质的翻译水平使B16细胞黑色素大量合成。%To study the effect of different media on the biological characteristics of B16 murine melanoma cells in vitro, and the expression mechanism of melanin, B16 cells were subcultured continuously in different media, and their biological characteristics including morphology, cell proliferation, cell cycle were observed. Meanwhile, the transcriptional and translational levels of melanin were investigated by Western Blot, RT-PCR, and two-dimensional electrophoresis. The results indicated that B16 murine melanoma cells had good cellular morphology, high cell viability, rapid proliferation, and could be subcultured continuously in two media, Gibco-RPMI-1640-31800 (RPMI-1640) and Gibco-DMEM-12800 (DMEM). However, melanin

  13. Carnosic acid inhibits the epithelial-mesenchymal transition in B16F10 melanoma cells: a possible mechanism for the inhibition of cell migration.

    Science.gov (United States)

    Park, So Young; Song, Hyerim; Sung, Mi-Kyung; Kang, Young-Hee; Lee, Ki Won; Park, Jung Han Yoon

    2014-01-01

    Carnosic acid is a natural benzenediol abietane diterpene found in rosemary and exhibits anti-inflammatory, antioxidant, and anti-carcinogenic activities. In this study, we evaluated the effects of carnosic acid on the metastatic characteristics of B16F10 melanoma cells. When B16F10 cells were cultured in an in vitro Transwell system, carnosic acid inhibited cell migration in a dose-dependent manner. Carnosic acid suppressed the adhesion of B16F10 cells, as well as the secretion of matrix metalloproteinase (MMP)-9, tissue inhibitor of metalloproteinase (TIMP)-1, urokinase plasminogen activator (uPA), and vascular cell adhesion molecule (VCAM)-1. Interestingly, secretion of TIMP-2 increased significantly in B16F10 cells treated with 10 μmol/L carnosic acid. Additionally, carnosic acid suppressed the mesenchymal markers snail, slug, vimentin, and N-cadherin and induced epithelial marker E-cadherin. Furthermore, carnosic acid suppressed phosphorylation of Src, FAK, and AKT. These results indicate that inhibition of the epithelial-mesenchymal transition may be important for the carnosic acid-induced inhibition of B16F10 cell migration. PMID:25036034

  14. Therapy of established B16-F10 melanoma tumors by a single vaccination of CTL/T helper peptides in VacciMax®

    Directory of Open Access Journals (Sweden)

    Korets-Smith Ella

    2007-04-01

    Full Text Available Abstract Background Melanoma tumors are known to express antigens that usually induce weak immune responses of short duration. Expression of both tumor-associated antigens p53 and TRP2 by melanoma cells raises the possibility of simultaneously targeting more than one antigen in a therapeutic vaccine. In this report, we show that VacciMax® (VM, a novel liposome-based vaccine delivery platform, can increase the immunogenicity of melanoma associated antigens, resulting in tumor elimination. Methods C57BL/6 mice bearing B16-F10 melanoma tumors were vaccinated subcutaneously 6 days post tumor implantation with a mixture of synthetic peptides (modified p53: 232–240, TRP-2: 181–188 and PADRE and CpG. Tumor growth was monitored and antigen-specific splenocyte responses were assayed by ELISPOT. Results Vaccine formulated in VM increased the number of both TRP2- and p53-specific IFN-γ producing splenocytes following a single vaccination. Vaccine formulated without VM resulted only in enhanced IFN-γ producing splenocytes to one CTL epitopes (TRP2:180–188, suggesting that VM overcomes antigen dominance and enhances immunogenicity of multiple epitopes. Vaccination of mice bearing 6-day old B16-F10 tumors with both TRP2 and p53-peptides formulated in VM successfully eradicated tumors in all mice. A control vaccine which contained all ingredients except liposomes resulted in eradication of tumors in no more than 20% of mice. Conclusion A single administration of VM is capable of inducing an effective CTL response to multiple tumor-associated antigens. The responses generated were able to reject 6-day old B16-F10 tumors.

  15. Bioactive Constituents of Zanthoxylum rhetsa Bark and Its Cytotoxic Potential against B16-F10 Melanoma Cancer and Normal Human Dermal Fibroblast (HDF) Cell Lines.

    Science.gov (United States)

    Santhanam, Ramesh Kumar; Ahmad, Syahida; Abas, Faridah; Safinar Ismail, Intan; Rukayadi, Yaya; Tayyab Akhtar, Muhammad; Shaari, Khozirah

    2016-01-01

    Zanthoxylum rhetsa is an aromatic tree, known vernacularly as "Indian Prickly Ash". It has been predominantly used by Indian tribes for the treatment of many infirmities like diabetes, inflammation, rheumatism, toothache and diarrhea. In this study, we identified major volatile constituents present in different solvent fractions of Z. rhetsa bark using GC-MS analysis and isolated two tetrahydrofuran lignans (yangambin and kobusin), a berberine alkaloid (columbamine) and a triterpenoid (lupeol) from the bioactive chloroform fraction. The solvent fractions and purified compounds were tested for their cytotoxic potential against human dermal fibroblasts (HDF) and mouse melanoma (B16-F10) cells, using the MTT assay. All the solvent fractions and purified compounds were found to be non-cytotoxic to HDF cells. However, the chloroform fraction and kobusin exhibited cytotoxic effect against B16-F10 melanoma cells. The presence of bioactive lignans and alkaloids were suggested to be responsible for the cytotoxic property of Z. rhetsa bark against B16-F10 cells. PMID:27231889

  16. Antitumor Effects In Vitro and In Vivo and Mechanisms of Protection against Melanoma B16F10-Nex2 Cells By Fastuosain, a Cysteine Proteinase from Bromelia fastuosa

    Directory of Open Access Journals (Sweden)

    Carla A. Guimarães-Ferreira

    2007-09-01

    Full Text Available In the present work, the antitumor effect of fastuosain, a cysteine proteinase from Bromelia fastuosa, was investigated. In the intravenous model of lung colonization in C57BI/6 mice, fastuosain and bromelain injected intraperitoneally were protective, very few nodules of B16F10-Nex2 melanoma cells were detected. Tumor cells treated with fastuosain showed reduced expression of CD44 and decreased invasion through Matrigel, lost their cytoplasmic extensions and substrate adherence, became round and detached, forming strongly bound cell clusters in suspension. Peritoneal cells recruited and activated by fastuosain treatment (mainly monocytic cells and lymphocytes migrated to the lung, where pulmonary melanoma metastases grew. Adoptive transference of peritoneal cells recruited by fastuosain had no protective effect against lung metastases in recipient mice. Treatment of green fluorescent protein -chimeric animals with fastuosain did not change the number of cells that migrated to the lung, compared to PBSinjected control mice, but the number of positive major histocompatibility complex class II cells increased with fastuosain treatment. Murine antibodies against fastuosain, bromelain, cathepsins B and L crossreacted in ELISA and recognized surface and cytoplasmic components expressed on B16F10-Nex2 cells. Anti-fastuosain antibodies were cytotoxic/lytic to B16F10-Nex2 cells. Antitumor effects of fastuosain involve mainly the direct effect of the enzyme and elicitation of protective antibodies.

  17. Antitumor effects in vitro and in vivo and mechanisms of protection against melanoma B16F10-Nex2 cells by fastuosain, a cysteine proteinase from Bromelia fastuosa.

    Science.gov (United States)

    Guimarães-Ferreira, Carla A; Rodrigues, Elaine G; Mortara, Renato A; Cabral, Hamilton; Serrano, Fabiana A; Ribeiro-dos-Santos, Ricardo; Travassos, Luiz R

    2007-09-01

    In the present work, the antitumor effect of fastuosain, a cysteine proteinase from Bromelia fastuosa, was investigated. In the intravenous model of lung colonization in C57Bl/6 mice, fastuosain and bromelain injected intraperitoneally were protective, and very few nodules of B16F10-Nex2 melanoma cells were detected. Tumor cells treated with fastuosain showed reduced expression of CD44 and decreased invasion through Matrigel, lost their cytoplasmic extensions and substrate adherence, and became round and detached, forming strongly bound cell clusters in suspension. Peritoneal cells recruited and activated by fastuosain treatment (mainly monocytic cells and lymphocytes) migrated to the lung, where pulmonary melanoma metastases grew. Adoptive transference of peritoneal cells recruited by fastuosain had no protective effect against lung metastases in recipient mice. Treatment of green fluorescent protein-chimeric animals with fastuosain did not change the number of cells that migrated to the lung, compared to PBS-injected control mice, but the number of positive major histocompatibility complex class II cells increased with fastuosain treatment. Murine antibodies against fastuosain, bromelain, and cathepsins B and L cross-reacted in ELISA and recognized surface and cytoplasmic components expressed on B16F10-Nex2 cells. Anti-fastuosain antibodies were cytotoxic/lytic to B16F10-Nex2 cells. Antitumor effects of fastuosain involve mainly the direct effect of the enzyme and elicitation of protective antibodies.

  18. Bioactive Constituents of Zanthoxylum rhetsa Bark and Its Cytotoxic Potential against B16-F10 Melanoma Cancer and Normal Human Dermal Fibroblast (HDF Cell Lines

    Directory of Open Access Journals (Sweden)

    Ramesh Kumar Santhanam

    2016-05-01

    Full Text Available Zanthoxylum rhetsa is an aromatic tree, known vernacularly as “Indian Prickly Ash”. It has been predominantly used by Indian tribes for the treatment of many infirmities like diabetes, inflammation, rheumatism, toothache and diarrhea. In this study, we identified major volatile constituents present in different solvent fractions of Z. rhetsa bark using GC-MS analysis and isolated two tetrahydrofuran lignans (yangambin and kobusin, a berberine alkaloid (columbamine and a triterpenoid (lupeol from the bioactive chloroform fraction. The solvent fractions and purified compounds were tested for their cytotoxic potential against human dermal fibroblasts (HDF and mouse melanoma (B16-F10 cells, using the MTT assay. All the solvent fractions and purified compounds were found to be non-cytotoxic to HDF cells. However, the chloroform fraction and kobusin exhibited cytotoxic effect against B16-F10 melanoma cells. The presence of bioactive lignans and alkaloids were suggested to be responsible for the cytotoxic property of Z. rhetsa bark against B16-F10 cells.

  19. Redox-Active Profile Characterization of Remirea maritima Extracts and It Cytotoxic Effect in Mouse Fibroblasts (L929 and Melanoma (B16F10 Cells

    Directory of Open Access Journals (Sweden)

    Grace Anne A. Dória

    2015-06-01

    Full Text Available Remirea maritima is a tropical plant with a reticulated root system belonging to the family Cyperaceae, also known to have biologically active secondary metabolites. However, very few data on R. maritima’s biological actions are available and there are no reports regarding the redox-active profile of this plant. In this study, we examined the total phenolic content of Remirea maritima hydroalcoholic (RMHA extracts, redox properties against different reactive species generated in vitro and their cytotoxic effect against fibroblasts (L929 and melanoma (B16F10 cells. Total reactive antioxidant potential index (TRAP and total antioxidant reactivity (TAR results revealed that RMHA at all concentrations tested showed significant antioxidant capacity. RMHA was also effective against hydroxyl radical formation, reduction of Fe3+ to Fe2+ and in scavenging nitric oxide (NO radicals. In vitro, the level of lipid peroxidation was reduced by RMHA extract and the data showed significant oxidative damage protection. The RMHA cytotoxicity was evaluated by a neutral red assay in fibroblast (L929 and melanome (B16F10 cells. The obtained results showed that the RMHA (40 and 80 µg/mL, respectively reduced 70% of the viable cells. In conclusion, this study represents the first report regarding the antioxidant and anti-proliferative potential of R. maritima against B16F10 melanoma cells.

  20. Antiproliferative activity and apoptosis induction of Eucalyptus Citriodora resin and its major bioactive compound in melanoma B16F10 cells.

    Science.gov (United States)

    Duh, Pin-Der; Chen, Zong-Tsi; Lee, Shwu-Woan; Lin, Tsuey-Pin; Wang, Ya-Ting; Yen, Wen-Jye; Kuo, Ling-Feng; Chu, Heuy-Ling

    2012-08-15

    Antiproliferative activity and apoptosis induction of ethyl acetate of Eucalyptus citriodora resin (EAEER), and its major bioactive compound in melanoma B16F10 cells were investigated. 6-[1-(p-Hydroxy-phenyl)ethyl]-7-O-methyl aromadendrin (HEMA), a flavanol derivative, was isolated from EAEER and identified on the basis of its mass and NMR spectra. The results from MTT assay showed high antiproliferative effects of EAEER and HEMA on B16F10 cells. Moreover, EAEER- and HEMA-induced cell apoptosis was association with the decrease in the mitochondrial transmembrane potentials (Δψ(m)), increase in Bax/Bcl-2 ratio, and activation of caspase-3. Cells treated with EAEER and HEMA generated intracellular reactive oxygen species (ROS) and nitric oxide (NO), indicating that ROS and RNS play important roles in the induction of apoptosis in B16F10 cells. Taken together, EAEER and its major bioactive compound, HEMA, inhibited the proliferation of B16F10 cells via apoptosis and may be a potential antimelanoma agent. PMID:22838509

  1. Establishment of Lymphangioma Model and a Study on the Promoting Effect of Murine Melanoma Cell B16-F1 on the Lymphangiogenesis In Vitro

    Institute of Scientific and Technical Information of China (English)

    CHEN Siyuau; CHEN Aijun; HUANG Chaugzheng; QIAN Yue; LIU Zhixiang; WU Yan; TU Yating

    2007-01-01

    To establish an animal model of benign lymphangiomas of C57BL/6 mouse in vitro and to observe the effect of mouse ascites melanoma cell B16-F1 on the lymphangiogenesis, 16 C57BL/6 mice aged 8 weeks were given two intraperitoneal injections of incomplete Freund's adjuvant at a15-day interval. The induced neoplasms were studied histopathologically and thhe neoplasms speci- mens were immunohistochemically examined for the expressions of VEGF-C (vascular endothelial growth factor-C) and Fit-4 (VEGFR-3, vascular endothelial growth factor receptor-3). The neoplasms were harvested and embedded in fibrin gel for culture in conditioned medium of B16-F1 cells in vitro and observed under inverted microscope. Our results showed that white solid tumor masses devel- oped in peritoneal cavity after the induction. The tumors were confirmed to be lymphangioma by gross and histological examination. The tumor cells expressed both VEGF-C and Flt-4. Lymphatic capillaries coming from lymphangioma specimen grew into the gel and the conditioned medium of B16-F1 cells was found to be able to promote the growth of the vessels. It is concluded that intrap- eritoneal injection of incomplete Freund's adjuvant is a good method for inducing benign lymphan- giomas in mouse and B16-F1 cells can promote lymphangiogenesis.

  2. Holothurian glycosaminoglycan inhibits metastasis and thrombosis via targeting of nuclear factor-κB/tissue factor/Factor Xa pathway in melanoma B16F10 cells.

    Directory of Open Access Journals (Sweden)

    Yang Zhao

    Full Text Available Holothurian glycosaminoglycan (hGAG is a high-molecular-weight form of fucosylated chondroitin sulfate and has an antithrombotic effect. Our previous studies demonstrated that hGAG efficiently inhibited tumor cell metastasis. The interplays between thrombosis and tumor progression may have a major impact on hematogenous metastasis. In this study, we demonstrated that the mouse melanoma B16F10 cells treated with hGAG displayed a significant reduction of metastasis and coagulation capacity in vitro and in vivo. Mechanistic studies revealed that hGAG treatment in B16F10 cells remarkably inhibited the formation of fibrin through attenuating the generation of activated Factor Xa (FXa, without affecting the expression of urokinase (uPA and plasminogen activator inhibitor 1 (PAI-1 that involved in fibrinolysis. Moreover, hGAG treatment downregulated the transcription and protein expression of tissue factor (TF. Promoter deletions, site mutations and functional studies identified that the nuclear transcription factor NF-κB binding region is responsible for hGAG-induced inhibition of TF expression. While the hGAG treatment of B16F10 cells was unable to inhibit NF-κB expression and phosphorylation, hGAG significantly prevented nuclear translocation of NF-κB from the cytosol, a potential mechanism underlying the transcriptional suppression of TF. Moreover, hGAG markedly suppressed the activation of p38MAPK and ERK1/2 signaling pathways, the central regulators for the expression of metastasis-related matrix metalloproteinases (MMPs. Consequently, hGAG exerts a dual function in the inhibition of metastasis and coagulation activity in mouse melanoma B16F10 cells. Our studies suggest hGAG to be a promising therapeutic agent for metastatic cancer treatment.

  3. Inhibition effect of phenyl compounds from the Oryza sativa roots on melanin production in murine B16-F10 melanoma cells.

    Science.gov (United States)

    Cho, Jin-Gyeong; Huh, Jeongran; Jeong, Rak-Hun; Cha, Byeong-Ju; Shrestha, Sabina; Lee, Dong-Geol; Kang, Hee-Cheol; Kim, Ji-Young; Baek, Nam-In

    2015-01-01

    Five phenyl compounds, vanillin (1), methyl trans-ferulate (2), trans-p-coumaric acid methyl ester (3), N-benzoyltryptamine (4), and N-(trans-cinnamoyl)tryptamine (5), were isolated from the roots of Oryza sativa L. and identified on the basis of spectroscopic data. Compounds 3 and 5 showed strong inhibition effect on melanin production in murine B16-F10 melanoma cells and tyrosinase activity. Also, the quantitative analysis of the compounds was carried out using LC/MS/MS experiment. Compounds 3 and 5 could be used as skin-whitening agents. PMID:25299734

  4. Paullinia cupana Mart var. sorbilis, guaraná, reduces cell proliferation and increases apoptosis of B16/F10 melanoma lung metastases in mice

    OpenAIRE

    Fukumasu, H.; J.L. Avanzo; Nagamine, M.K.; J.A. Barbuto; Rao, K.V.; M.L.Z. Dagli

    2008-01-01

    We showed that guaraná (Paullinia cupana Mart var. sorbilis) had a chemopreventive effect on mouse hepatocarcinogenesis and reduced diethylnitrosamine-induced DNA damage. In the present experiment, we evaluated the effects of guaraná in an experimental metastasis model. Cultured B16/F10 melanoma cells (5 x 10(5) cells/animal) were injected into the tail vein of mice on the 7th day of guaraná treatment (2.0 mg P. cupana/g body weight, per gavage) and the animals were treated with guaraná daily...

  5. Identification of donor melanoma in a renal transplant recipient.

    Science.gov (United States)

    Wilson, L J; Horvat, R T; Tilzer, L; Meis, A M; Montag, L; Huntrakoon, M

    1992-12-01

    A patient with chronic renal failure received a closely matched cadaveric kidney. Approximately 3 months after transplantation, the patient developed a metastatic malignant melanoma. A large retroperitoneal mass consisting of large pleomorphic polygonal neoplastic cells was found close to the donated kidney. This tumor was diagnosed as a malignant melanoma. DNA analysis of this tumor, the donated kidney, and the recipient indicated that the melanoma originated from the donor. Although this is not the first report of a donated melanoma, it is the first report of definitive DNA analysis of the origin of the malignant cells.

  6. Electrotransfer of Plasmid DNA Encoding an Anti-Mouse Endoglin (CD105 shRNA to B16 Melanoma Tumors with Low and High Metastatic Potential Results in Pronounced Anti-Tumor Effects

    Directory of Open Access Journals (Sweden)

    Tanja Dolinsek

    2015-12-01

    Full Text Available Endoglin overexpression is associated with highly proliferative tumor endothelium and also with some tumors, including melanoma. Its targeting has anti-tumor effectiveness, which can also be obtained by RNA interference. The aim of our study was to explore the anti-tumor effectiveness of endoglin silencing by electrotransfer of plasmid DNA encoding short hairpin RNA against endoglin in two murine B16 melanoma variants with different metastatic potential on cells, spheroids and subcutaneous tumors in mice. The results demonstrate that endoglin silencing with gene electrotransfer reduces the proliferation, survival and migration of melanoma cells and also has anti-tumor effectiveness, as the therapy resulted in a high percentage of tumor cures (23% and 58% on B16F1 and B16F10 tumors, respectively. The effectiveness of the therapy correlated with endoglin expression in melanoma cells; in vitro the effects were more pronounced in B16F1 cells, which express more endoglin than B16F10. However, the opposite was observed in vivo in tumors, where there was a higher expression of endoglin and better anti-tumor effectiveness in the B16F10 tumor. In conclusion, targeting endoglin for the treatment of melanoma seems to be a concept worthy of further exploration due to the increased therapeutic effect of the therapy based on simultaneous vascular targeting and its direct effect on tumor cells.

  7. Comparative Depigmentation Effects of Resveratrol and Its Two Methyl Analogues in α-Melanocyte Stimulating Hormone-Triggered B16/F10 Murine Melanoma Cells.

    Science.gov (United States)

    Yoon, Hoon-Seok; Hyun, Chang-Gu; Lee, Nam-Ho; Park, Sung-Soo; Shin, Dong-Bum

    2016-06-01

    Previous research showed that resveratrol (trans-3,4',5-trihydroxystilbene) and pinostilbene (trans-3-methoxy-4',5-dihydroxystilbene) were able to inhibit tyrosinase directly; however, anti-melanogenic effects of pterostilbene (trans-3,5-dimethoxy-4'-hydroxystilbene) and resveratrol trimethyl ether (RTE) have not been compared. To investigate the hypopigmentation effects of pterostilbene and RTE, melanin contents and intracellular tyrosinase activity were determined by western blot analysis. Firstly, pterostilbene showed the inhibitory effects on α-melanocyte stimulating hormone (MSH)-induced melanin synthesis stronger than RTE, resveratrol, and arbutin. Pterostilbene inhibited melanin biosynthesis in a dose-dependent manner in α-MSH-stimulated B16/F10 murine melanoma cells. Specifically, melanin content and intracellular tyrosinase activity were inhibited by 63% and 58%, respectively, in response to treatment with 10 μM of pterostilbene. The results of western blot analysis indicated that pterostilbene induced downregulation of tyrosinase protein expression and suppression of α-MSH-stimulated melan-A protein expression stronger than RTE or resveratrol. Based on these results, our study suggests that pterostilbene can induce hypopigmentation effects more effectively than resveratrol and RTE, and it functions via downregulation of protein expression associated with hyperpigmentation in α-MSH-triggered B16/F10 murine melanoma cells. PMID:27390733

  8. Antiangiogenesis, Loss of Cell Adhesion and Apoptosis Are Involved in the Antitumoral Activity of Proteases from V. cundinamarcensis (C. candamarcensis in Murine Melanoma B16F1

    Directory of Open Access Journals (Sweden)

    Dalton Dittz

    2015-03-01

    Full Text Available The proteolytic enzymes from V. cundinamarcensis latex, (P1G10, display healing activity in animal models following various types of lesions. P1G10 or the purified isoforms act as mitogens on fibroblast and epithelial cells by stimulating angiogenesis and wound healing in gastric and cutaneous ulcers models. Based on evidence that plant proteinases act as antitumorals, we verified this effect on a murine melanoma model. The antitumoral effect analyzed mice survival and tumor development after subcutaneous administration of P1G10 into C57BL/6J mice bearing B16F1 low metastatic melanoma. Possible factors involved in the antitumoral action were assessed, i.e., cytotoxicity, cell adhesion and apoptosis in vitro, haemoglobin (Hb, vascular endothelial growth factor (VEGF, tumor growth factor-β (TGF-β, tumor necrosis factor-α (TNF-α content and N-acetyl-glucosaminidase (NAG activity. We observed that P1G10 inhibited angiogenesis measured by the decline of Hb and VEGF within the tumor, and TGF-β displayed a non-significant increase and TNF-α showed a minor non-significant reduction. On the other hand, there was an increase in NAG activity. In treated B16F1 cells, apoptosis was induced along with decreased cell binding to extracellular matrix components (ECM and anchorage, without impairing viability.

  9. Comparative Depigmentation Effects of Resveratrol and Its Two Methyl Analogues in α-Melanocyte Stimulating Hormone-Triggered B16/F10 Murine Melanoma Cells

    Science.gov (United States)

    Yoon, Hoon-Seok; Hyun, Chang-Gu; Lee, Nam-Ho; Park, Sung-Soo; Shin, Dong-Bum

    2016-01-01

    Previous research showed that resveratrol (trans-3,4′,5-trihydroxystilbene) and pinostilbene (trans-3-methoxy-4′,5-dihydroxystilbene) were able to inhibit tyrosinase directly; however, anti-melanogenic effects of pterostilbene (trans-3,5-dimethoxy-4′-hydroxystilbene) and resveratrol trimethyl ether (RTE) have not been compared. To investigate the hypopigmentation effects of pterostilbene and RTE, melanin contents and intracellular tyrosinase activity were determined by western blot analysis. Firstly, pterostilbene showed the inhibitory effects on α-melanocyte stimulating hormone (MSH)-induced melanin synthesis stronger than RTE, resveratrol, and arbutin. Pterostilbene inhibited melanin biosynthesis in a dose-dependent manner in α-MSH-stimulated B16/F10 murine melanoma cells. Specifically, melanin content and intracellular tyrosinase activity were inhibited by 63% and 58%, respectively, in response to treatment with 10 μM of pterostilbene. The results of western blot analysis indicated that pterostilbene induced downregulation of tyrosinase protein expression and suppression of α-MSH-stimulated melan-A protein expression stronger than RTE or resveratrol. Based on these results, our study suggests that pterostilbene can induce hypopigmentation effects more effectively than resveratrol and RTE, and it functions via downregulation of protein expression associated with hyperpigmentation in α-MSH-triggered B16/F10 murine melanoma cells. PMID:27390733

  10. Melanogenesis stimulation in B16-F10 melanoma cells induces cell cycle alterations, increased ROS levels and a differential expression of proteins as revealed by proteomic analysis

    Energy Technology Data Exchange (ETDEWEB)

    Cunha, Elizabeth S.; Kawahara, Rebeca [Departamento de Bioquimica e Biologia Molecular, Setor de Ciencias Biologicas, Universidade Federal do Parana, P.O. Box 19046, CEP 81531-990, Curitiba, PR (Brazil); Kadowaki, Marina K. [Universidade Estadual do Oeste do Parana, Cascavel, PR (Brazil); Amstalden, Hudson G.; Noleto, Guilhermina R.; Cadena, Silvia Maria S.C.; Winnischofer, Sheila M.B. [Departamento de Bioquimica e Biologia Molecular, Setor de Ciencias Biologicas, Universidade Federal do Parana, P.O. Box 19046, CEP 81531-990, Curitiba, PR (Brazil); Martinez, Glaucia R., E-mail: grmartinez@ufpr.br [Departamento de Bioquimica e Biologia Molecular, Setor de Ciencias Biologicas, Universidade Federal do Parana, P.O. Box 19046, CEP 81531-990, Curitiba, PR (Brazil)

    2012-09-10

    Considering that stimulation of melanogenesis may lead to alterations of cellular responses, besides melanin production, our main goal was to study the cellular effects of melanogenesis stimulation of B16-F10 melanoma cells. Our results show increased levels of the reactive oxygen species after 15 h of melanogenesis stimulation. Following 48 h of melanogenesis stimulation, proliferation was inhibited (by induction of cell cycle arrest in the G1 phase) and the expression levels of p21 mRNA were increased. In addition, melanogenesis stimulation did not induce cellular senescence. Proteomic analysis demonstrated the involvement of proteins from other pathways besides those related to the cell cycle, including protein disulfide isomerase A3, heat-shock protein 70, and fructose biphosphate aldolase A (all up-regulated), and lactate dehydrogenase (down-regulated). In RT-qPCR experiments, the levels of pyruvate kinase M2 mRNA dropped, whereas the levels of ATP synthase (beta-F1) mRNA increased. These data indicate that melanogenesis stimulation of B16-F10 cells leads to alterations in metabolism and cell cycle progression that may contribute to an induction of cell quiescence, which may provide a mechanism of resistance against cellular injury promoted by melanin synthesis. -- Highlights: Black-Right-Pointing-Pointer Melanogenesis stimulation by L-tyrosine+NH{sub 4}Cl in B16-F10 melanoma cells increases ROS levels. Black-Right-Pointing-Pointer Melanogenesis inhibits cell proliferation, and induced cell cycle arrest in the G1 phase. Black-Right-Pointing-Pointer Proteomic analysis showed alterations in proteins of the cell cycle and glucose metabolism. Black-Right-Pointing-Pointer RT-qPCR analysis confirmed alterations of metabolic targets after melanogenesis stimulation.

  11. ADHESION-INDUCE PROTEIN TYROSINE PHOSPHORY-LATION IS ASSOCIATED WITH INVASIVE AND METASTATIC POTENTIALS IN B16-BL6 MELANOMA CELLS

    Institute of Scientific and Technical Information of China (English)

    Yan Chunhong; Han Rui

    1998-01-01

    Objective: The interaction of cancer cell with extracellular matrix (ECM) happens as an earlier and specific event in the invasive and metastatic cascade. To explore the key element(s) in cancer metastasis and observe the cell-ECM interaction and its role. Methods:To interrupt the cell-ECM interaction by suppression of adhesion-induced protein tyrosine phosphorylation with protein tyrosine kinase inhibitor genistein in B16-B16mouse melanoma cells. Results: When B16-BL6 cells attached to Matrigel, a solubilized basement membrane preparation from EHS sarcoma, a 125 kDa protein increased its phosphotyrosine content dramatically. In contrast, when the cells were pretreated with 20μM or 30μM genistein for 3 days, it was revealed a less increase in the phosphotyrosine content of this 125 kDa protein inresponse to cell attachment to ECM was revealed with immunoblot analysis. Accompanied by the lower level of adhesion-induced protein tyrosine phosphorylation the genistein-treated cells exhibited a decrease in their capabilities of adhesion to Matrigel and invasion through reconstituted basement membrane. The potentials of and forming lung metastatic nodules were also shown to be decreased dramatically in these genistein-treated cells.Conclusion: It was suggested that protein tyrosine phosphorylation in cell-ECM interaction might be associated with invasive and metastatic potentials in cancer cells.

  12. Transmission of donor melanoma to multiple organ transplant recipients.

    Science.gov (United States)

    Morris-Stiff, G; Steel, A; Savage, P; Devlin, J; Griffiths, D; Portman, B; Mason, M; Jurewicz, W A

    2004-03-01

    Malignant melanoma represents the most common tumour responsible for donor-derived post transplantation malignancies. We report the varied presentation and outcome of three graft recipients (two kidney and hepatic) who developed metastatic melanoma following cadaveric organ transplantation from a single multiorgan donor. Two of the recipients presented with symptomatic metastatic lesions and the third patient, despite being carefully monitored, developed evidence of metastatic cutaneous melanoma. Two of the patients died as a direct result of their melanomas. The recipients of corneal and cardiac grafts remain disease-free. We conclude that despite careful screening, donor-derived tumours remain a not uncommon clinical entity. The identification of a lesion in one recipient should prompt immediate examination and investigation of the remaining recipients of multiorgan donations.

  13. Inhibitory Effect of Dried Pomegranate Concentration Powder on Melanogenesis in B16F10 Melanoma Cells; Involvement of p38 and PKA Signaling Pathways

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    Su Jin Kang

    2015-10-01

    Full Text Available Plants rich in antioxidant substances may be useful for preventing skin aging. Pomegranates, containing flavonoids and other polyphenolic compounds, are widely consumed due to their beneficial properties. We examined the underlying mechanisms of dried pomegranate concentrate powder (PCP on melanin synthesis in B16F10 melanoma cells. The antioxidant effects of PCP were determined by measuring free radical scavenging capacity and transcript levels of antioxidant enzymes. To explore the inhibitory effects of PCP on melanin synthesis, we measured tyrosinase activity and melanin content in α-melanocyte stimulating hormone (α-MSH-stimulated B16F10 cells. In addition, the levels of tyrosinase-related protein-1 (TRP-1, TRP-2, tyrosinase, and microphthalmia-associated transcription factor (MITF expression were determined by Western blotting. Changes in the phosphorylation status of protein kinase A (PKA, cAMP response element-binding protein (CREB, mitogen-activated protein kinases (MAPKs, phosphatidylinositol 3-kinase (PI3K, serine/threonine kinase Akt, and glycogen kinase 3β (GSK3β were also examined. The free radical scavenging activity of PCP increased in a dose-dependent manner. In PCP-treated B16F10 cells, transcript levels of glutathione peroxidase-1 (GPx-1 were increased compared with α-MSH-stimulated cells. In addition, PCP led to the down-regulation of phospho-p38, phospho-PKA, phospho-CREB, phospho-GSK3β, MITF, and TRP-1 compared with α-MSH-stimulated B16F10 cells. We believe this effect may be associated with PCP activity, which leads to the inhibition of melanin production and tyrosinase activity. These results suggest that PCP decreases tyrosinase activity and melanin production via inactivation of the p38 and PKA signaling pathways, and subsequently decreases phosphorylation of CREB, MITF, and melanogenic enzymes. These observations provided new insights on the molecular mechanisms of the skin-whitening property of PCP.

  14. Cell cycle regulation by the Wee1 Inhibitor PD0166285, Pyrido [2,3-d] pyimidine, in the B16 mouse melanoma cell line

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    Sakamoto Masaharu

    2006-12-01

    Full Text Available Abstract Background Wee1 kinase plays a critical role in maintaining G2 arrest through its inhibitory phosphorylation of cdc2. In previous reports, a pyridopyrimidine molecule PD0166285 was identified to inhibit Wee1 activity at nanomolar concentrations. This G2 checkpoint abrogation by PD0166285 was demonstrated to kill cancer cells, there at a toxic highest dose of 0.5 μM in some cell lines for exposure periods of no longer than 6 hours. The deregulated cell cycle progression may have ultimately damaged the cancer cells. We herein report one of the mechanism by which PD0166285 leads to cell death in the B16 mouse melanoma cell line. Methods Tumor cell proliferation was determined by counting cell numbers. Cell cycle distribution was determined by flow cytometry. Morphogenesis analysis such as microtubule stabilization, Wee1 distribution, and cyclin B location were observed by immunofluorescence confocal microscopy. An immunoblot analysis of cdc2-Tyr15, cyclin D, E, p16, 21, 27, and Rb. A real-time PCR of the mRNA of cyclin D were completed. Results In our experiment, B16 cells also dramatically abrogated the G2 checkpoint and were found to arrest in the early G1 phase by treatment with 0.5 μM for 4 hours observed by flow cytometry. Cyclin D mRNA decreased within 4 hours observed by Real-time PCR. Rb was dephosphrylated for 24 hours. However, B16 cells did not undergo cell death after 0.5 μM treatment for 24 hours. Immnofluoscence microscopy showed that the cells become round and small in the morphogenesis. More interesting phenomena were that microtubule stabilization was blocked, and Wee1 distribution was restricted after treatment for 4 hours. Conclusion We analyzed the effect of Wee1 inhibitor PD0166285 described first by Wang in the G2 transition in the B16 melanoma cell line. The inhibitor PD0166285 abrogated G2/M checkpoint inducing early cell division. Moreover, we found that the treatment of cells with the inhibitor is related to

  15. Inhibitory Effects of Adlay Extract on Melanin Production and Cellular Oxygen Stress in B16F10 Melanoma Cells

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    Huey-Chun Huang

    2014-09-01

    Full Text Available The aim of this study was to determine the effects of adlay extract on melanin production and the antioxidant characteristics of the extract. The seeds were extracted by the supercritical fluid CO2 extraction (SFE method. The effect of adlay extract on melanin production was evaluated using mushroom tyrosinase activity assay, intracellular tyrosinase activity, antioxidant properties and melanin content. Those assays were performed spectrophotometrically. In addition, the expression of melanogenesis-related proteins was determined by western blotting. The results revealed that the adlay extract suppressed intracellular tyrosinase activity and decreased the amount of melanin in B16F10 cells. The adlay extract decreased the expression of microphthalmia-associated transcription factor (MITF, tyrosinase, tyrosinase related protein-1 (TRP-1 and tyrosinase related protein-2 (TRP-2. The extract also exhibited antioxidant characteristics such as free radical scavenging capacity and reducing power. It effectively decreased intracellular reactive oxygen species (ROS levels in B16F10 cells. We concluded that the adlay extract inhibits melanin production by down-regulation of MITF, tyrosinase, TRP-1 and TRP-2. The antioxidant properties of the extract may also contribute to the inhibition of melanogenesis. The adlay extract can therefore be applied as an inhibitor of melanogenesis and could also act as a natural antioxidant in skin care products.

  16. Inhibitory effect of Cucumis sativus on melanin production in melanoma B16 cells by downregulation of tyrosinase expression.

    Science.gov (United States)

    Kai, Hisahiro; Baba, Masaki; Okuyama, Toru

    2008-12-01

    We compared the inhibitory effects on melanogenesis of six plant parts (leaves, stems, roots, whole fruits, calyxes, and fruits without calyxes) of Cucumis sativus. MeOH extracts of leaves and stems inhibited melanin production in B16 cells. These extracts did not affect the activity of mushroom tyrosinase or crude enzyme lysate from B16 cells. However, the extracts decreased tyrosinase expression at the protein level. These results suggest that the depigmenting mechanism of extracts from leaves and stems of C. SATIVUS involves the expression of tyrosinase. Of eight compounds isolated from the leaves, lutein ( 1) (IC (50) = 170.7 microM) and (+)-(1 R,2 S,5 R,6 S)-2,6-di-(4'-hydroxyphenyl)-3,7-dioxabicyclo[3.3.0]octane ( 2) (IC (50) = 270.8 microM) were found to suppress melanogenesis. Whereas 1 was found to markedly decrease the expression levels of tyrosinase, 2 only weakly reduced tyrosinase expression. This suggests that 1 is an active component in the leaves of C. sativus and is a potentially useful skin-whitening agent. PMID:19009501

  17. The leaf extract ofMallotus japonicus and its major active constituent, rutin, suppressed on melanin production in murine B16F1 melanoma

    Institute of Scientific and Technical Information of China (English)

    Junsei Taira; Eito Tsuchida; Masatsugu Uehara; Natsuko Ohhama; Wakana Ohmine; Takayuki Ogi

    2015-01-01

    Objective:To find anti-melanogenesis materials used in whitening cosmetics. Methods: The ethanolic leaf extract ofMallotus japonicus (M. japonicus) having an anti-melanogenesis activity was separated by a sephadexLH-20 chromatography. Each fraction was measured for its tyrosinase inhibitory activity together with its polyphenol content using the Folin–Ciocalteu method. The anti-melanogenesis activity of the active fractions was determined by the melanin content in the murine B16F1 melanoma. The active fractions were put together due to similar constituents, and then separated by high performance liquid chromatography using a C-18 ODS column. The major anti-melanogenesis compound was identified using 1Hand13C-NMR and liquid chromatography-mass spectrometry. Results: The ethanolic leaf extract ofM. japonicus showed an anti-tyrosinase activity with a high polyphenol content, resulting in suppression of melanin production in the B16F1 melanoma. The extract was separated and the active compound was identical as rutin based on the1H,13C-NMR and liquid chromatography-mass spectrometry analysis data. In addition, the rutin treatment with cells reduced the melanin content in a concentration dependent manner without any cell toxicity. The leaf extract ofM. japonicus containing rutin would be useful in whitening cosmetics for protection from UV-light exposure to be limiting the accumulation of melanin in skin. Conclusions: The leaf extract ofM. japonicus and/or rutin isolated from the extract as a key whitening agent would be useful as a whitening cosmetic material for protecting against disorder skin due to melanin accumulation.

  18. Tumor-targeted delivery of a C-terminally truncated FADD (N-FADD) significantly suppresses the B16F10 melanoma via enhancing apoptosis

    Science.gov (United States)

    Yang, Yun-Wen; Zhang, Chun-Mei; Huang, Xian-Jie; Zhang, Xiao-Xin; Zhang, Lin-Kai; Li, Jia-Huang; Hua, Zi-Chun

    2016-01-01

    Fas-associated protein with death domain (FADD), a pivotal adaptor protein transmitting apoptotic signals, is indispensable for the induction of extrinsic apoptosis. However, overexpression of FADD can form large, filamentous aggregates, termed death effector filaments (DEFs) by self-association and initiate apoptosis independent of receptor cross-linking. A mutant of FADD, which is truncated of the C-terminal tail (m-FADD, 182–205 aa) named N-FADD (m-FADD, 1–181 aa), can dramatically up-regulate the strength of FADD self-association and increase apoptosis. In this study, it was found that over-expression of FADD or N-FADD caused apoptosis of B16F10 cells in vitro, even more, N-FADD showed a more potent apoptotic effect than FADD. Meanwhile, Attenuated Salmonella Typhimurium strain VNP20009 was engineered to express FADD or N-FADD under the control of a hypoxia-induced NirB promoter and each named VNP-pN-FADD and VNP-pN-N-FADD. The results showed both VNP-pN-FADD and VNP-pN-N-FADD delayed tumor growth in B16F10 mice model, while VNP-pN-N-FADD suppressed melanoma growth more significantly than VNP-pN-FADD. Additionally, VNP-pN-FADD and VNP-pN-N-FADD induced apoptosis of tumor cells by activating caspase-dependent apoptotic pathway. Our results show that N-FADD is a more potent apoptotic inducer and VNP20009-mediated targeted expression of N-FADD provides a possible cancer gene therapeutic approach for the treatment of melanoma. PMID:27767039

  19. Activation of MITF by Argan Oil Leads to the Inhibition of the Tyrosinase and Dopachrome Tautomerase Expressions in B16 Murine Melanoma Cells.

    Science.gov (United States)

    Villareal, Myra O; Kume, Sayuri; Bourhim, Thouria; Bakhtaoui, Fatima Zahra; Kashiwagi, Kenichi; Han, Junkyu; Gadhi, Chemseddoha; Isoda, Hiroko

    2013-01-01

    Argan (Argania spinosa L.) oil has been used for centuries in Morocco as cosmetic oil to maintain a fair complexion and to cure skin pimples and chicken pox pustules scars. Although it is popular, the scientific basis for its effect on the skin has not yet been established. Here, the melanogenesis regulatory effect of argan oil was evaluated using B16 murine melanoma cells. Results of melanin assay using B16 cells treated with different concentrations of argan oil showed a dose-dependent decrease in melanin content. Western blot results showed that the expression levels of tyrosinase (TYR), tyrosinase-related protein 1 (TRP1), and dopachrome tautomerase (DCT) proteins were decreased. In addition, there was an increase in the activation of MITF and ERK1/2. Real-time PCR results revealed a downregulation of Tyr, Trp1, Dct, and Mitf mRNA expressions. Argan oil treatment causes MITF phosphorylation which subsequently inhibited the transcription of melanogenic enzymes, TYR and DCT. The inhibitory effect of argan oil on melanin biosynthesis may be attributed to tocopherols as well as the synergistic effect of its components. The results of this study provide the scientific basis for the traditionally established benefits of argan oil and present its therapeutic potential against hyperpigmentation disorders.

  20. Activation of MITF by Argan Oil Leads to the Inhibition of the Tyrosinase and Dopachrome Tautomerase Expressions in B16 Murine Melanoma Cells

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    Myra O. Villareal

    2013-01-01

    Full Text Available Argan (Argania spinosa L. oil has been used for centuries in Morocco as cosmetic oil to maintain a fair complexion and to cure skin pimples and chicken pox pustules scars. Although it is popular, the scientific basis for its effect on the skin has not yet been established. Here, the melanogenesis regulatory effect of argan oil was evaluated using B16 murine melanoma cells. Results of melanin assay using B16 cells treated with different concentrations of argan oil showed a dose-dependent decrease in melanin content. Western blot results showed that the expression levels of tyrosinase (TYR, tyrosinase-related protein 1 (TRP1, and dopachrome tautomerase (DCT proteins were decreased. In addition, there was an increase in the activation of MITF and ERK1/2. Real-time PCR results revealed a downregulation of Tyr, Trp1, Dct, and Mitf mRNA expressions. Argan oil treatment causes MITF phosphorylation which subsequently inhibited the transcription of melanogenic enzymes, TYR and DCT. The inhibitory effect of argan oil on melanin biosynthesis may be attributed to tocopherols as well as the synergistic effect of its components. The results of this study provide the scientific basis for the traditionally established benefits of argan oil and present its therapeutic potential against hyperpigmentation disorders.

  1. Activation of MITF by Argan Oil Leads to the Inhibition of the Tyrosinase and Dopachrome Tautomerase Expressions in B16 Murine Melanoma Cells.

    Science.gov (United States)

    Villareal, Myra O; Kume, Sayuri; Bourhim, Thouria; Bakhtaoui, Fatima Zahra; Kashiwagi, Kenichi; Han, Junkyu; Gadhi, Chemseddoha; Isoda, Hiroko

    2013-01-01

    Argan (Argania spinosa L.) oil has been used for centuries in Morocco as cosmetic oil to maintain a fair complexion and to cure skin pimples and chicken pox pustules scars. Although it is popular, the scientific basis for its effect on the skin has not yet been established. Here, the melanogenesis regulatory effect of argan oil was evaluated using B16 murine melanoma cells. Results of melanin assay using B16 cells treated with different concentrations of argan oil showed a dose-dependent decrease in melanin content. Western blot results showed that the expression levels of tyrosinase (TYR), tyrosinase-related protein 1 (TRP1), and dopachrome tautomerase (DCT) proteins were decreased. In addition, there was an increase in the activation of MITF and ERK1/2. Real-time PCR results revealed a downregulation of Tyr, Trp1, Dct, and Mitf mRNA expressions. Argan oil treatment causes MITF phosphorylation which subsequently inhibited the transcription of melanogenic enzymes, TYR and DCT. The inhibitory effect of argan oil on melanin biosynthesis may be attributed to tocopherols as well as the synergistic effect of its components. The results of this study provide the scientific basis for the traditionally established benefits of argan oil and present its therapeutic potential against hyperpigmentation disorders. PMID:23935660

  2. Maslinic Acid, a Triterpene from Olive, Affects the Antioxidant and Mitochondrial Status of B16F10 Melanoma Cells Grown under Stressful Conditions

    Science.gov (United States)

    Mokhtari, Khalida; Rufino-Palomares, Eva E.; Pérez-Jiménez, Amalia; Reyes-Zurita, Fernando J.; Figuera, Celeny; García-Salguero, Leticia; Medina, Pedro P.; Peragón, Juan; Lupiáñez, José A.

    2015-01-01

    Maslinic acid (MA) is a natural compound whose structure corresponds to a pentacyclic triterpene. It is abundant in the cuticular lipid layer of olives. MA has many biological and therapeutic properties related to health, including antitumor, anti-inflammatory, antimicrobial, antiparasitic, antihypertensive, and antioxidant activities. However, no studies have been performed to understand the molecular mechanism induced by this compound in melanoma cancer. The objective of this study was to examine the effect of MA in melanoma (B16F10) cells grown in the presence or absence of fetal bovine serum (FBS). We performed cell proliferation measurements, and the reactive oxygen species (ROS) measurements using dihydrorhodamine 123 (DHR 123) and activities of catalase, glucose 6-phosphate dehydrogenase, glutathione S-transferase, and superoxide dismutase. These changes were corroborated by expression assays. FBS absence reduced cell viability decreasing IC50 values of MA. The DHR 123 data showed an increase in the ROS level in the absence of FBS. Furthermore, MA had an antioxidant effect at lower assayed levels measured as DHR and antioxidant defense. However, at higher dosages MA induced cellular damage by apoptosis as seen in the results obtained. PMID:26236377

  3. Holothurian glycosaminoglycan inhibits metastasis via inhibition of P-selectin in B16F10 melanoma cells.

    Science.gov (United States)

    Yue, Zhiqiang; Wang, Aiyun; Zhu, Zhijie; Tao, Li; Li, Yao; Zhou, Liang; Chen, Wenxing; Lu, Yin

    2015-12-01

    P-selectin-mediated tumor cell adhesion to platelets is a well-established stage in the process of tumor metastasis. Through computerized structural analysis, we found a marine-derived polysaccharide, holothurian glycosaminoglycan (hGAG), behaved as a ligand-competitive inhibitor of P-selectin, indicating its potential to disrupt the binding of P-selectin to cell surface receptor and activation of downstream regulators of tumor cell migration. Our experimental data demonstrated that hGAG significantly inhibited P-selectin-mediated adhesion of tumor cells to platelets and tumor cell migration in vitro and reduced subsequent pulmonary metastasis in vivo. Furthermore, abrogation of the P-selectin-mediated adhesion of tumor cells led to down-regulation of protein levels of integrins, FAK and MMP-2/9 in B16F10 cells, which is a crucial molecular mechanism of hGAG to inhibit tumor metastasis. In conclusion, hGAG has emerged as a novel anti-cancer agent via blocking P-selectin-mediated malignant events of tumor metastasis.

  4. The Cytolytic Amphipathic β(2,2-Amino Acid LTX-401 Induces DAMP Release in Melanoma Cells and Causes Complete Regression of B16 Melanoma.

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    Liv-Marie Eike

    Full Text Available In the present study we examined the ability of the amino acid derivative LTX-401 to induce cell death in cancer cell lines, as well as the capacity to induce regression in a murine melanoma model. Mode of action studies in vitro revealed lytic cell death and release of danger-associated molecular pattern molecules, preceded by massive cytoplasmic vacuolization and compromised lysosomes in treated cells. The use of a murine melanoma model demonstrated that the majority of animals treated with intratumoural injections of LTX-401 showed complete and long-lasting remission. Taken together, these results demonstrate the potential of LTX-401 as an immunotherapeutic agent for the treatment of solid tumors.

  5. Paullinia cupana Mart var. sorbilis, guaraná, reduces cell proliferation and increases apoptosis of B16/F10 melanoma lung metastases in mice

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    H. Fukumasu

    2008-04-01

    Full Text Available We showed that guaraná (Paullinia cupana Mart var. sorbilis had a chemopreventive effect on mouse hepatocarcinogenesis and reduced diethylnitrosamine-induced DNA damage. In the present experiment, we evaluated the effects of guaraná in an experimental metastasis model. Cultured B16/F10 melanoma cells (5 x 10(5 cells/animal were injected into the tail vein of mice on the 7th day of guaraná treatment (2.0 mg P. cupana/g body weight, per gavage and the animals were treated with guaraná daily up to 14 days until euthanasia (total treatment time: 21 days. Lung sections were obtained for morphometric analysis, apoptotic bodies were counted to calculate the apoptotic index and proliferating cell nuclear antigen-positive cells were counted to determine the proliferation index. Guaraná-treated (GUA animals presented a 68.6% reduction in tumor burden area compared to control (CO animals which were not treated with guaraná (CO: 0.84 ± 0.26, N = 6; GUA: 0.27 ± 0.24, N = 6; P = 0.0043, a 57.9% reduction in tumor proliferation index (CO: 23.75 ± 20.54, N = 6; GUA: 9.99 ± 3.93, N = 6; P = 0.026 and a 4.85-fold increase in apoptotic index (CO: 66.95 ± 22.95, N = 6; GUA: 324.37 ± 266.74 AB/mm², N = 6; P = 0.0152. In this mouse model, guaraná treatment decreased proliferation and increased apoptosis of tumor cells, consequently reducing the tumor burden area. We are currently investigating the molecular pathways of the effects of guaraná in cultured melanoma cells, regarding principally the cell cycle inhibitors and cyclins.

  6. Inhibition of honokiol on the proliferation and melanin biosynthesis on B16 melanoma cells in vitro%和厚朴酚对小鼠黑色素瘤B16细胞增殖以及黑色素合成的影响

    Institute of Scientific and Technical Information of China (English)

    喻丽红; 张超; 谭茵

    2012-01-01

    Objective To investigate the inhibition of honokiol on proliferation and melanin biosynthesis in B16 melanoma cells in vitro. Methods B16 cells were cultured in vitro. MTT assay, DAPI and NaOH lysis test were applied for assessment of cell proliferation, cellular morphologic observation and cellular melanin content, respectively. Results B16 cells were treated with honokiol with different concentrations ( 5,10,20,40 and 80 μmol/L ) and durations ( 12,24 and 48 h ). The IC50 of honokiol on proliferation of B16 cells were 23. 4, 13. 1 and 11.4 μmol/L, respectively, with durations of 12,24 and 48 h. Apoptosis of B16 cells were induced with honokiol in DAPI staining. Furthermore, the melanin biosynthesis was inhibited with honokiol with concentration over 20 (xmol/L. Conclusion Honokiol effectively inhibits the proliferation of B16 in dose - and time - depend manners. Apoptotic bodies were observed in B16 cells treated with honokiol. Although melanin biosynthesis is also inhibited by honokiol, this effect is insignificant.%目的 探讨和厚朴酚对小鼠黑色素瘤B16细胞增殖以及细胞内黑色素合成的影响.方法 体外培养小鼠B16细胞,MTT法检测和厚朴酚对B16细胞增殖的影响;DAPI染色法观察和厚朴酚对B16细胞细胞形态的影响;NaOH裂解法检测和厚朴酚对黑色素含量的影响.结果 和厚朴酚浓度为5、10、20、40、80 μmol/L 作用B16细胞,在不同的用药时间12、24和48 h,和厚朴酚对B16细胞增殖的IC50分别为23.4、13.1和11.4 μmol/L;同时和厚朴酚作用B16细胞12 h后,经DAPI染色,B16细胞呈现典型的凋亡形态;另外采用20、40、80 μmol/L的和厚朴酚分别作用B16细胞,B16细胞内黑色素合成量也呈现降低的趋势.结论 和厚朴酚能有效地抑制B16细胞增殖,且其抑制作用具有时间和浓度依赖性;药物作用后的B16细胞呈现凋亡形态并出现凋亡小体;和厚朴酚对B16细胞内黑色素合成也呈现抑制作用但不明显.

  7. Antitumor Effects In Vitro and In Vivo and Mechanisms of Protection against Melanoma B16F10-Nex2 Cells By Fastuosain, a Cysteine Proteinase from Bromelia fastuosa1

    OpenAIRE

    Guimarães-Ferreira, Carla A; Rodrigues, Elaine G; Renato A Mortara; Cabral, Hamilton; Fabiana A. Serrano; Ribeiro-dos-Santos, Ricardo; Travassos, Luiz R.

    2007-01-01

    In the present work, the antitumor effect of fastuosain, a cysteine proteinase from Bromelia fastuosa, was investigated. In the intravenous model of lung colonization in C57Bl/6 mice, fastuosain and bromelain injected intraperitoneally were protective, and very few nodules of B16F10-Nex2 melanoma cells were detected. Tumor cells treated with fastuosain showed reduced expression of CD44 and decreased invasion through Matrigel, lost their cytoplasmic extensions and substrate adherence, and beca...

  8. Effects of serum containing Tribulus terrestris on proliferation and melanogenesis in cultured B16 melanoma cells%刺蒺藜血清对小鼠B16黑素瘤细胞增殖及黑素合成的影响

    Institute of Scientific and Technical Information of China (English)

    杨柳; 谢家宝; 洪逊强; 刘利利; 马玲玲

    2011-01-01

    Objective To observe the effects of serum containing Tribulus terrestris on proliferation and melanogenesis in B16 melanoma cells.Methods B16 melanoma cells was cultured in vitro.Serum containing Tribulus terrestris was prepared.MTT and NaOH method were used to detect proliferation and melanogenesis respectively.Results The serum containing Tribulus terrestris increased the amount of melanin at higher concentrations (30g) but acted to the reverse effect at lower concentrations (10g) (P<0.01).Compared with blank control group,The amount of melanin at middle concentrations (20g) had no significant difference (P>0.05).Conclusions the effects of serum containing Tribulus terrestris on proliferation and melanogenesis in B16 melanoma cells correlated with the concentration of Tribulus terrestris.lt presented a two-way regulation on proliferation and melanogenesis in cultured B16 melanoma cells.%目的:探讨刺蒺藜血清对黑素瘤细胞增殖及黑素合成的影响.方法:体外培养小鼠B16黑素瘤细胞,制备大鼠含药血清,分别采用四甲基偶氮唑蓝比色法(MTT法)和NaOH裂解法检测细胞增殖及黑素含量.结果:刺蒺藜对黑素细胞增殖及黑素含量具有低剂量(10g)抑制、高剂量(30g)促进作用(P<0.01),中剂量(20g)对细胞增殖及黑素含量的影响倾向于促进作用,但与空白对照组比较无统计学意义.结论:刺蒺藜对黑素细胞增殖及黑素含量的影响与剂量有关,具有低浓度抑制,高浓度激活的双向调节作用.

  9. In vitro and in vivo antitumor efficacy of CLA-PTX on B16-F10 melanoma cells%CLA-PTX对黑色素瘤B16-F10的体内外抗肿瘤作用

    Institute of Scientific and Technical Information of China (English)

    李捷思; 杨科; 柯曦宇; 杜若; 张烜; 张强

    2014-01-01

    本研究旨在探索CLA-PTX对黑色素瘤B16-F10细胞的体内外抗肿瘤作用.选择鼠源B16-F10细胞系作为研究模型.研究CLA-PTX体外细胞毒,细胞凋亡,细胞周期作用,以及CLA-PTX体外细胞摄取作用.用荷瘤C57BL6/N小鼠研究CLA-PTX的体内抗肿瘤作用.体外细胞毒研究结果表明:CLA-PTX的ICso为(4.25±0.43) μM,优于紫杉醇(6.70±0.80) μM(P<0.01).与空白组和紫杉醇组相比,CLA-PTX可使细胞总凋亡比例增加(P<0.01).与空白对照组相比,CLA-PTX将细胞周期阻滞于S期,而紫杉醇则使细胞在G2-M期蓄积.CLA-PTX的细胞摄取量显著高于紫杉醇(P<0.01).体内抗肿瘤药效显示,CLA-PTX抗肿瘤活性显著高于空白对照组和紫杉醇组(P<0.01或P<0.05).上述结果表明,CLA-PTX对B16-F10细胞有显著体内外抗肿瘤作用.

  10. Differentiation-inducing and anti-proliferative activities of isoliquiritigenin and all-trans-retinoic acid on B16F0 melanoma cells: Mechanisms profiling by RNA-seq.

    Science.gov (United States)

    Chen, Xiaoyu; Yang, Ming; Hao, Wenjin; Han, Jichun; Ma, Jun; Wang, Caixia; Sun, Shiguo; Zheng, Qiusheng

    2016-10-30

    Melanoma is a cancer that arises from melanocytes, specialized pigmented cells that are found predominantly in the skin. The incidence of malignant melanoma has significantly increased over the last decade. With the development of therapy, the survival rate of some kind of cancer has been improved greatly. But the treatment of melanoma remains unsatisfactory. Much of melanoma's resistance to traditional chemotherapy is believed to arise intrinsically, by virtue of potent growth and cell survival-promoting genetic alteration. Therefore, significant attention has recently been focused on differentiation therapy, as well as differentiation inducer compounds. In previous study, we found isoliquiritigenin (ISL), a natural product extracted from licorice, could induce B16F0 melanoma cell differentiation. Here we investigated the transcriptional response of melanoma differentiation process induced by ISL and all-trans-retinoic acid (RA). Results showed that 390 genes involves in 201 biochemical pathways were differentially expressed in ISL treatment and 304 genes in 193 pathways in RA treatment. Differential expressed genes (DGEs, fold-change (FC)≥10) with the function of anti-proliferative and differentiation inducing indicated a loss of grade malignancy characteristic. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated glutathione metabolism, glycolysis/gluconeogenesis and pentose phosphate pathway were the top three relative pathway perturbed by ISL, and mitogen-activated protein kinase (MAPK) signaling pathway was the most important pathway in RA treatment. In the analysis of hierarchical clustering of DEGs, we discovered 72 DEGs involved in the process of drug action. We thought Cited1, Tgm2, Xaf1, Cd59a, Fbxo2, Adh7 may have critical role in the differentiation of melanoma. The evidence displayed herein confirms the critical role of reactive oxygen species (ROS) in melanoma pathobiology and provides evidence for future targets in the

  11. Anti-Tumor Effects of B16F10 Tumor Cell Lysate Vaccine in Mouse Melanoma%肿瘤细胞裂解物抗小鼠B16F10黑色素瘤的作用研究

    Institute of Scientific and Technical Information of China (English)

    路蕾; 刘景晶; 陈科; 刘彬; 王泽宇; 葛驰宇; 侯景; 金亮; 邢芸; 曹荣月

    2012-01-01

    反复冻融B16F10肿瘤细胞制备裂解物,以白喉毒素(Diphtheria toxin,DT)为载体,OK432和来源于结核分枝杆菌( Mycobacterium tuberculosis)热休克蛋白70(HSP70)第407-426( mHSP70407-426,M)的两段串联重复序列M2为佐剂,制备了肿瘤细胞疫苗B16F10-DT-M2-OK432(BDTMOK),探讨其能否抑制小鼠B16黑色素瘤,并且对其抗肿瘤的作用机理进行部分探讨.以制备的BDTMOK免疫C57BL/6小鼠,分别检测体液免疫应答和细胞免疫应答.通过ELISA法,从血清中检测到高滴度的抗B16肿瘤细胞裂解物(B16 tumor cell lysate,B16TCL)类抗体.淋巴细胞增殖实验的结果显示,BDTMOK的免疫能够有效的刺激脾淋巴细胞的增殖.预防结合治疗性实验的结果显示,BDTMOK激发的免疫应答对于B16肿瘤攻击起到有效的保护作用,与PBS阴性对照组比较,皮下注射BDTMOK可以延长皮下移植瘤发生的潜伏期(P<0.05),并且平均瘤重显著降低(P<0.05);抑制了小鼠皮内肿瘤模型中的血管新生(P<0.01).疫苗BDTMOK能有效的抑制小鼠B16黑色素瘤的生长.%To make tumor vaccine B16F10-DT-M2-OK432,the freeze-thaw method was adopted to obtain B16F10 tumor cell lysate. Diphtheria toxin( DT) ,OK432 or M2 which was two tandem repeats of sequence 407-426 of microbial HSP70 were selected as carrier and adjuvants respectively. The C57BL/6 mice were immunized with B16F10-DT-M2-OK432 in order to explore whether the tumor cell vaccine can effectively inhibit B16F10 melanoma in tumor-bearing mice and to study the mechanism of B16F10-DT-M2-OK432. After the last immunization, humoral immune and cellular immune response were detected. High tiler of anti-B16F10 tumor cell lysate antibody was detected in immunized mice sera by ELISA. Splenic lymophocyte proliferation assay results showed that the proliferation activity of splenocytes from mice immunized with B16F10-DT-M2-OK432 vaccine was significantly higher. The results of prophylactic/ therapeutic experiment

  12. Antitumor Effects In Vitro and In Vivo and Mechanisms of Protection against Melanoma B16F10-Nex2 Cells By Fastuosain, a Cysteine Proteinase from Bromelia fastuosa1

    Science.gov (United States)

    Guimarães-Ferreira, Carla A; Rodrigues, Elaine G; Mortara, Renato A; Cabral, Hamilton; Serrano, Fabiana A; Ribeiro-dos-Santos, Ricardo; Travassos, Luiz R

    2007-01-01

    In the present work, the antitumor effect of fastuosain, a cysteine proteinase from Bromelia fastuosa, was investigated. In the intravenous model of lung colonization in C57Bl/6 mice, fastuosain and bromelain injected intraperitoneally were protective, and very few nodules of B16F10-Nex2 melanoma cells were detected. Tumor cells treated with fastuosain showed reduced expression of CD44 and decreased invasion through Matrigel, lost their cytoplasmic extensions and substrate adherence, and became round and detached, forming strongly bound cell clusters in suspension. Peritoneal cells recruited and activated by fastuosain treatment (mainly monocytic cells and lymphocytes) migrated to the lung, where pulmonary melanoma metastases grew. Adoptive transference of peritoneal cells recruited by fastuosain had no protective effect against lung metastases in recipient mice. Treatment of green fluorescent protein-chimeric animals with fastuosain did not change the number of cells that migrated to the lung, compared to PBS-injected control mice, but the number of positive major histocompatibility complex class II cells increased with fastuosain treatment. Murine antibodies against fastuosain, bromelain, and cathepsins B and L cross-reacted in ELISA and recognized surface and cytoplasmic components expressed on B16F10-Nex2 cells. Anti-fastuosain antibodies were cytotoxic/lytic to B16F10-Nex2 cells. Antitumor effects of fastuosain involve mainly the direct effect of the enzyme and elicitation of protective antibodies. PMID:17898868

  13. Acetylsalicylic acid regulates MMP-2 activity and inhibits colorectal invasion of murine B16F0 melanoma cells in C57BL/6J mice: effects of prostaglandin F(2)alpha.

    Science.gov (United States)

    Tsai, Chin-Shaw Stella; Luo, Shue-Fen; Ning, Chung-Chu; Lin, Chien-Liang; Jiang, Ming-Chung; Liao, Ching-Fong

    2009-08-01

    Epidemiological studies indicate that acetylsalicylic acid may reduce the risk of mortality due to colon cancers. Metastasis is the major cause of cancer death. Matrix metalloproteinases (MMPs) play important roles in tumor invasion regulation, and prostaglandin F(2)alpha (PGF(2)alpha) is a key stimulator of MMP production. Thus, we investigated whether acetylsalicylic acid regulated MMP activity and the invasion of cancer cells and whether PGF(2)alpha attenuated acetylsalicylic acid-inhibited invasion of cancer cells. Gelatin-based zymography assays showed that acetylsalicylic acid inhibited the MMP-2 activity of B16F0 melanoma cells. Matrigel-based chemoinvasion assays showed that acetylsalicylic acid inhibited the invasion of B16F0 cells. Acetylsalicylic acid can inhibit PGF(2)alpha synthesis and PGF(2)alpha is a key stimulator of MMP-2 production. Our data showed that PGF(2)alpha treatment attenuated the acetylsalicylic acid-inhibited invasion of B16F0 cells. In animal experiments, acetylsalicylic acid reduced colorectal metastasis of B16F0 cells in C57BL/6J mice by 44%. Our results suggest that PGF(2)alpha is a therapeutic target for metastasis inhibition and acetylsalicylic acid may possess anti-metastasis ability.

  14. 多次高温热疗对小鼠B16黑色素瘤疗效的实验研究%Anticancer Effect of B16 Malignant Melanoma in C57BL/6 Mice by Multiple Hyperthermia

    Institute of Scientific and Technical Information of China (English)

    徐国权; 赵凌云; 王晓文; 刘继光; 唐劲天

    2009-01-01

    目的 本研究探索多次加温治疗对小鼠B16黑色素瘤疗效的相关治疗机制,比较加温次数对恶性黑色素瘤疗效的影响.方法 实验采用自行开发的加热加湿生物实验平台,通过比较小鼠生存期,肿瘤重量,观察各组肿瘤的组织形态学变化,以及利用流式细胞术检测小鼠外周血中CD4+、CD8+、CD28+的表达来评价肿瘤治疗的效果.结果 实验中各热疗组小鼠的生存时间要长于对照组(P<0.05),且以热疗3次组的小鼠生存期最长.各热疗组的肿瘤生长明显受到抑制(P<0.05),且热疗3次组的抑瘤效果最明显.组织病理学观察发现热疗组有大量肿瘤细胞呈凋亡、坏死样改变,热疗3次组瘤旁组织中有大量淋巴细胞浸润.此外同对照组比较各热疗组CD4+、CD8+、CD28+的表达均有升高(P<0.05).结论 多次热疗抑制肿瘤生长和激发机体免疫的效果要优于单次热疗.

  15. Non-melanoma skin cancer in Portuguese kidney transplant recipients - incidence and risk factors*

    Science.gov (United States)

    Pinho, André; Gouveia, Miguel; Cardoso, José Carlos; Xavier, Maria Manuel; Vieira, Ricardo; Alves, Rui

    2016-01-01

    Background Cancer is currently among the three leading causes of death after solid organ transplantation and its incidence is increasing. Non-melanoma skin cancer - squamous cell carcinoma and basal cell carcinoma - is the most common malignancy found in kidney transplant recipients (KTRs). The incidence of non-melanoma skin cancer in KTRs has not been extensively studied in Portugal. Objectives To determine the incidence of non-melanoma skin cancer in KTRs from the largest Portuguese kidney transplant unit; and to study risk factors for non-melanoma skin cancer. Methods Retrospective analysis of clinical records of KTRs referred for the first time for a dermatology consultation between 2004 and 2013. A case-control study was performed on KTRs with and without non-melanoma skin cancer. Results We included 288 KTRs with a median age at transplantation of 47 years, a male gender predominance (66%) and a median transplant duration of 3.67 years. One fourth (n=71) of KTRs developed 131 non-melanoma skin cancers, including 69 (53%) squamous cell carcinomas and 62 (47%) basal cell carcinomas (ratio squamous cell carcinoma: basal cell carcinoma 1.11), with a mean of 1.85 neoplasms per patient. Forty percent of invasive squamous cell carcinomas involved at least two clinical or histological high-risk features. The following factors were associated with a higher risk of non-melanoma skin cancer: an older age at transplantation and at the first consultation, a longer transplant duration and the presence of actinic keratosis. KTRs treated with azathioprine were 2.85 times more likely to develop non-melanoma skin cancer (p=0.01). Conclusion Non-melanoma skin cancer was a common reason for dermatology consultation in Portuguese KTRs. It is imperative for KTRs to have access to specialized dermatology consultation for early referral and treatment of skin malignancies. PMID:27579740

  16. Irradiated survival effect of melanoma B16 cells at different penetration depths of intermediate energy heavy-ion beams%中能碳离子束对不同贯穿深度上黑色素瘤B16细胞的辐照存活效应

    Institute of Scientific and Technical Information of China (English)

    李强; 周光明; 王菊芳; 李文建; 温小琼; 党秉荣; 颉红梅; 卫增泉

    2001-01-01

    Survival fractions of melanoma B16 cells at different penetrationdepths exposed to 49.30MeV/u 12C ion beam and modulated 74.55MeV/u 12C ion beam with a ridge filter to form a spread-out Bragg peak (SOBP) were measured. The results show the survival response of B16 cells as the biological endpoint to heavy ions increases synchronously with the deposited energy along the beam traversal, that is, the survival fraction near or at the Bragg peak or SOBP is far lower than that at the beam's entrance or exit channel. Comparisons of the experimental data with calculated survival fractions at different depths predicted by the linear-quadratic survival model combined with dose-averaged LET display a good agreement within the experimental errors. It is prompted that the linear-quadratic model combined with dose-averaged LET possibly is a valid way to calculate the survival effect of B16 cells along the beam penetration under the exposure of the intermediate energy heavy-ion beam.%测量了49.30MeV/u及经脊形过滤器展宽的Bragg峰的74.55MeV/u12C离子束在不同贯穿深度上辐射后黑色素瘤B16细胞的存活率。结果显示,以存活为终点的B16细胞生物学效应与碳离子束能量沉积同步增大,即Bragg或展宽的Bragg峰区内细胞存活率远低于离子束入射或出射通道处的细胞存活率。通过实验测量与剂量平均LET结合的线性平方模型理论计算存活曲线的比较,发现在实验测量误差范围内两者符合较好。提示:剂量平均LET结合线性平方模型可能是预测中能重离子束贯穿深度上B16细胞存活效应的一种有效的方法。

  17. Regression of subcutaneous B16 melanoma tumors after intratumoral delivery of an IL-15-expressing plasmid followed by in vivo electroporation

    OpenAIRE

    Ugen, KE; Kutzler, MA; Marrero, B; Westover, J; Coppola, D; Weiner, DB; Heller, R.

    2006-01-01

    In vivo electroporation has been used to efficiently deliver drugs and ‘therapeutic’ genes to tumors, including melanoma lesions. This study reports on the effect of intratumoral delivery of an optimized DNA plasmid expressing interleukin-15 (pIL-15) on established murine melanoma tumors. IL-15 has been demonstrated to have a pivotal role in the function of memory CD8+ T cells and natural killer cells, which are critical for tumor immunosurveillance. In this study, C57BL/6 mice were injected ...

  18. BFD-22 a new potential inhibitor of BRAF inhibits the metastasis of B16F10 melanoma cells and simultaneously increased the tumor immunogenicity

    NARCIS (Netherlands)

    Ferreira, Adilson Kleber; Mesquita Pasqualoto, Kerly Fernanda; Kruyt, Frank A. E.; Palace-Berl, Fanny; Azevedo, Ricardo Alexandre; Turra, Kely Medeiros; Rodrigues, Cecilia Pessoa; Franco Ferreira, Ana Carolina; Clavijo Salomon, Maria Alejandra; de Sa Junior, Paulo Luiz; Farias, Camyla Fernandes; Figueiredo, Carlos Rogerio; Tavares, Leoberto Costa; Marzagdo Barbuto, Jose Alexandre; Jorge, Salomao Doria

    2016-01-01

    Benzofuroxan is an interesting ring system, which has shown a wide spectrum of biological responses against tumor cell lines. We investigated, herein, the antitumor effects of benzofuroxan derivatives (BFDs) in vitro and in a melanoma mouse model. Cytotoxic effects of twenty-two BFDs were determined

  19. 转染人核糖核酸酶抑制因子基因对B16黑色瘤细胞及肿瘤转移的影响%Effects of Transfection of Human Ribonuclease Inhibitor Gene on B16 Melanoma Cells and Tumor Metastasis

    Institute of Scientific and Technical Information of China (English)

    陈俊霞; 田余祥; 傅攀峰; 夏俊; 阎平; 崔秀云

    2004-01-01

    Human ribonuclease inhibitor (RI) is an acidic cytoplasmic glycoprotein with molecular mass of 50 ku. RI can inhibit the activity of ribonuclease A ( RNase A). Angiogenin (Ang) is a member of RNase A superfamily. RI also can inhibit Ang activities by tight combination. Angiogenesis is an essential condition for the development of tumors and their metastatic dissemination. So anti-angiogenesis will be an efficient method in the inhibition of the growth and metastasis of tumor. The experiment demonstrated that RI might effectively block the angiogenesis that was induced by angiogenin. RI is constructed almost entirely of leucine-rich repeats that might involve in other unclear biological effect. In order to understand further the potential function of RI and investigate the role of RI in invasion and metastasis. The study established a transfection of human RI cDNA into B16 melanoma cells by the retroviral packaging cell line PA317 carrying the pLNCX-RI in vitro. Transfected B16 cells by PA317 carrying the pLNCX and untransfected B16 cells were used as control. The B16pLNCX-RI cell line with a stably high expression of RI was identified by PCR, RT-PCR, Western blot and immunofluorescence assay respectively. The results showed that the transfected RI gene might significantly inhibit cell proliferation, migration, and enhance cell adhesion, as well as, make morphological changes in vitro. Cell doubling time were (24.98 ±0.16) h, (25.62 ±0.28) h, (32.64 + 1.11) h in B16 cells, B16 pLNCX and B16 pLNCX-RI cells respectively. Cell adhesion rate was significantly increased by 19. 5% and 17. 8% as well as cell migration was reduced by 60% and 61.4% in B16 pLNCX-RI cells compared with pLNCX B16 cells and B16 cells respectively. B16 pLNCX-RI cells became flatter, less nucleoli, less division phases and weaker alkalophilic quality of cytoplasm compared with control groups, which should imply that cell proliferation viability was decreased and malignant phenotype was improved

  20. Bio-efficacy of Tea Extract on Mouse B16 Melanoma Cell in Vitro and Its Mechanism%茶叶提取物对B16小鼠黑色素瘤细胞的生物学效应及机理研究

    Institute of Scientific and Technical Information of China (English)

    陈贞纯; 贾玲燕; 毛祖法; 金恩惠; 王广伟; 屠幼英

    2014-01-01

    研究了茶叶提取物对体外培养的 B16小鼠黑色素瘤细胞的生物学效应,并对其机理进行初步探讨。分别用茶黄素单体(TF1)、EGCG、纯度为40%的茶黄素(TF40)处理细胞,然后观察细胞形态,并以溴化二苯四偶氮法(MTT 法)测定受试物对黑色素细胞活力的影响,以左旋多巴为底物测定细胞内酪氨酸酶活性,采用比色法测定细胞中的黑色素含量。实验结果表明,各化合物对B16黑色素瘤细胞形态有不同程度的影响,对细胞活性、酪氨酸酶浓度和黑色素合成量有浓度依赖性抑制作用。这3种茶提取物能通过多种途径抑制黑色素合成,从而达到美白的效果,且茶叶提取物的效果优于Vc。%The Bio-efficacy and mechanism of tea extract on mouse B16 melanoma cell in vitro were studied in this paper. Cells were treated with different concentration of TF1, EGCG and TF40, then the cell morphology was observed. The tyrosinase activity was determined by MTT method. The synthesis of melanin was determined using DOPA as substrate, and proliferation rate of B16 cells was determined by colorimetry, then analyzing the effects of drugs to these indexes. The results showed that the cell morphology was changed by the treatment of drugs. The tyrosinase activity, the synthesis of melanin and proliferation rate of B16 cells were all negatively related to the drugs concentration. Tea extracts and Vc can cause the inhibition of the synthesis of melanin, and result ed to white skin. The whiting effect of tea extracts is better than that of Vc.

  1. 6种中药组方对小鼠B16黑素瘤细胞增殖和黑素生成的影响%The Effect of Six Kinds of Traditional Chinese Herbal Composing Prescriptions on Cell Proliferation and Melanogenesis in Cultured B16 Melanoma Cells

    Institute of Scientific and Technical Information of China (English)

    张建; 巫玮; 万玉华; 张榕文; 刘丹丹; 麻睿; 顾为望

    2011-01-01

    目的 研究6种中药组方水提物、醇提物对小鼠B16黑素瘤细胞增殖及黑素生成的影响.方法 以MTF法测定各组方对B16细胞的增殖活性:NaOH裂解法测定各组方对B16细胞的黑素生成量.结果 组方1、2、3水提物和组方2、4、6醇提物具有明显促细胞增殖作用(P>0.05);水提物中细胞增殖率最高的为组方2(0.625 mg/ml),增殖率为22.60%;醇提物中细胞增殖率最高的为组方4(0.078 mg/ml),增殖率为11.60%.组方1、2、4、5、6水提物和6种组方醇提物均具有促进黑色素生成的作用(P<0.05).水提物中黑素增率最高的为组方1(0.313 mg/ml),增率为75.38%;醇提物中黑素生成增率最高的为组方1(1.25 mg/ml),增率为88.01%.结论 各组方对鼠黑素瘤B16细胞增殖及黑素生成均有一定的促进作用,但其作用强度与组方的组成密切相关.%Objective To study the effects of six ethanol extracts and aqueous extracts from traditional Chinese herbal composing prescriptions on the cell proliferation and melanogenesis in cultured B 16 melanoma cells. Methods The cell proliferation were measured by MTT method, and NaOH assay was to determine melanin synthesis. Results The aqueous extracts of composition 1,2,3 and alcohol extracts of composition 2,4,6 had significantly promoted cell proliferation (P<0.05); aqueous extract with the highest rate of cell proliferation was composing prescription 2 (0.625 mg/ml), the proliferation rate was 22.60%; ethanol extract with the highest rate of cell proliferation was composition 4 (0.078 mg/ml),the proliferation rate was 11.60%. The aqueous extract of composing prescriptions 1,2,4,5,6 and alcohol extracts of the six kinds of composing prescriptions promoted obviously the synthesis of melanin (P<0.05).Aqueous extract with the highest rate of melanin synthesis was composing prescription 1 (0.313 mg/ml),the growth rate is 75.38%; alcohol extract with the highest rate of melanin synthesis was composing

  2. Accumulation of cytolytic CD8{sup +} T cells in B16-melanoma and proliferation of mature T cells in TIS21-knockout mice after T cell receptor stimulation

    Energy Technology Data Exchange (ETDEWEB)

    Ryu, Min Sook [Department of Biochemistry and Molecular Biology, Ajou University School of Medicine, 164, World cul-ro, Yeongtong-gu, Suwon, Gyeonggi-do 443-380 (Korea, Republic of); Woo, Min-Yeong [Department of Microbiology, Ajou University School of Medicine, 164, World cul-ro, Yeongtong-gu, Suwon, Gyeonggi-do 443-380 (Korea, Republic of); Department of Biomedical Sciences, The Graduate School, Ajou University (Korea, Republic of); Kwon, Daeho [Department of Microbiology, Kwandong University College of Medicine, Gangneung, Gangwon-do 210-701 (Korea, Republic of); Hong, Allen E. [Department of Biochemistry and Molecular Biology, Ajou University School of Medicine, 164, World cul-ro, Yeongtong-gu, Suwon, Gyeonggi-do 443-380 (Korea, Republic of); Song, Kye Yong [Department of Pathology, Chung-Ang University College of Medicine, Dongjak-gu, Seoul 156-756 (Korea, Republic of); Park, Sun [Department of Microbiology, Ajou University School of Medicine, 164, World cul-ro, Yeongtong-gu, Suwon, Gyeonggi-do 443-380 (Korea, Republic of); Lim, In Kyoung [Department of Biochemistry and Molecular Biology, Ajou University School of Medicine, 164, World cul-ro, Yeongtong-gu, Suwon, Gyeonggi-do 443-380 (Korea, Republic of)

    2014-10-01

    In vivo and in vitro effects of TIS21 gene on the mature T cell activation and antitumor activities were explored by employing MO5 melanoma orthograft and splenocytes isolated from the TIS21-knockout (KO) mice. Proliferation and survival of mature T cells were significantly increased in the KO than the wild type (WT) cells, indicating that TIS21 inhibits the rate of mature T cell proliferation and its survival. In MO5 melanoma orthograft model, the KO mice recruited much more CD8{sup +} T cells into the tumors at around day 14 after tumor cell injection along with reduced tumor volumes compared with the WT. The increased frequency of granzyme B{sup +} CD8{sup +} T cells in splenocytes of the KO mice compared with the WT may account for antitumor-immunity of TIS21 gene in the melanoma orthograft. In contrast, reduced frequencies of CD107a{sup +} CD8{sup +} T cells in the splenocytes of KO mice may affect the loss of CD8{sup +} T cell infiltration in the orthograft at around day 19. These results indicate that TIS21 exhibits antiproliferative and proapoptotic effects in mature T cells, and differentially affects the frequencies of granzyme B{sup +} CD8{sup +} T-cells and CD107a{sup +} CD8{sup +} T-cells, thus transiently regulating in vivo anti-tumor immunity. - Highlights: • Constitutive expression of TIS21 in splenocytes and upregulation by TCR stimulation. • Proliferation of mature T-cells in spleen of TIS21KO mice after TCR stimulation. • Inhibition of cell death in mature T-cells of TIS21KO mice compared with the wild type. • Inhibition of melanoma growth in TIS21KO mice and CD8{sup +} T cell infiltration in tumor. • Reduction of CD 107{sup +}CD8{sup +} T cells, but increased granzyme B{sup +} CD8{sup +} T cells in TIS21KO mice.

  3. Over-expression of ornithine decarboxylase antizyme fusion proteins affects the cell cycle of mouse melanoma B16-F1 cells%鸟氨酸脱羧酶抗酶融合蛋白高表达对小鼠黑素瘤细胞B16-F1细胞周期的影响

    Institute of Scientific and Technical Information of China (English)

    刘梦瑶; 韩钰; 蔡富强; 何玲; 王艳林

    2011-01-01

    This study aims to assess the effects of over-expressed GFP-0AZ1 (green fluorescent protein-or-nithine decarboxylase antizyme-1) and GFP-OAZ2 (green fluorescent protein-omithine decarboxylase antizyme-2) fusion proteins on cell cycle of mouse melanoma B16-F1 cells. GFP-OAZ1 and GFP-OAZ2 fusion genes were constructed truly, then transiently transfected into B16-F1 cells by lipofectamine reagent. The expressions of GFP-OAZ1 and GFP-OAZ2 fusion proteins were confirmed by immunocytochemistry and Western blot analysis. Flow cytometry was applied to detect the effect of the fusion proteins on the cell cycle of B16-F1 cells; Western blot analysis was also used to detect the effect of GFP-0AZ1/2 on ornithine decarboxylase (ODC) in protein level. Results showed that over-expression of GFP-OAZ1 and GFP-0AZ2 fusion proteins in B16-F1 resulted in G1/G0 arrest in the cell cycle. When OAZ1 or OAZ2 gene was co-transfected with ODC gene into B16-F1 cells, over-expression of GFP-OAZ1 fusion protein, not GFP-OAZ2, demonstrated the ability to significantly decrease the total protein level of ODC. We concluded that over-expression of GFP-OAZ1 or GFP-OAZ2 fusion gene could lead to cell cycle arrest in Gl/GO phase, and GFP-0AZ1 fusion protein could stimulate ODC degradation efficiently, but no such function was found on OAZ2 fusion protein.%研究鸟氨酸脱羧酶抗酶(OAZ)与绿色荧光蛋白融合蛋白GFP-OAZ1和GFP-OAZ2高表达对小鼠黑色素瘤B16-F1细胞周期的影响.构建GFP-OAZ1和GFP-OAZ2融合基因的真核表达质粒并经脂质体法瞬时转染B16-F1细胞,然后用Western blot分析法和荧光显微镜下观察融合蛋白在B16-F1细胞中的表达.流式细胞分析用于检测GFP-OAZ融合蛋白高表达对B16-F1细胞周期的影响.Western blot分析法鉴定GFP-OAZ融合蛋白高表达对B16-F1细胞中鸟氨酸脱羧酶(ODC)酶蛋白水平的影响.结果成功构建的GFP-OAZ1和GFP-OAZ2融合基因B16-F1细胞中正确高效表达.

  4. Synthesis of O-(2-[18F]fluoroethyl)-L-tyrosine and its biological evaluation in B16 melanoma-bearing mice as PET tracer for tumor imaging

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    O-(2-[18F]fluoroethyl) -L-tyrosine([18F]FET) ,a fluorine-18 labeled analogue of tyrosine,has been syn-thesized and biologically evaluated in tumor-bearing mice. The whole synthesis procedure is com-pleted within 50 min. The radiochemical yield is about 40%(no decay corrected) and radiochemical purity more than 97% after simplified solid phase extraction. [18F]FET shows rapid,high uptake and long retention in the tumor as well as low uptake in the brain. The ratios of tumor-to-muscle(T/M) and tumor-to-blood(T/B) of [18F]FET are similar to those of [18F]FDG,but the ratios of tumor-to-brain(T/Br) are 2-3 times higher than that of [18F]FDG. Autoradiography of [18F]FET demonstrates a remarkable accumulation in melanoma with high contrast. It appears to be a probable competitive candidate for melanoma imaging with PET.

  5. Synthesis of O-(2-[18F]fluoroethyl)-L-tyrosine and its biological evaluation in B16 melanoma-bearing mice as PET tracer for tumor imaging

    Institute of Scientific and Technical Information of China (English)

    WANG MingWei; YIN DuanZhi; LI ShiQiang; WANG YongXian

    2007-01-01

    O-(2-[18F]fluoroethyl)-L-tyrosine ([18F]FET), a fluorine-18 labeled analogue of tyrosine, has been synthesized and biologically evaluated in tumor-bearing mice. The whole synthesis procedure is completed within 50 min. The radiochemical yield is about 40% (no decay corrected) and radiochemical purity more than 97% after simplified solid phase extraction. [18F]FET shows rapid, high uptake and long retention in the tumor as well as low uptake in the brain. The ratios of tumor-to-muscle (T/M) and tumor-to-blood (T/B) of [18F]FET are similar to those of [18F]FDG, but the ratios of tumor-to-brain (T/Br)are 2-3 times higher than that of [18F]FDG. Autoradiography of [18F]FET demonstrates a remarkable accumulation in melanoma with high contrast. It appears to be a probable competitive candidate for melanoma imaging with PET.

  6. Cross-talk between 5-hydroxytryptamine and substance P in the melanogensis and apoptosis of B16F10 melanoma cells.

    Science.gov (United States)

    Zhou, Jia; Geng, Kun-kun; Ping, Feng-feng; Gao, Yue-ying; Liu, Lei; Feng, Bai-nian

    2016-03-15

    Skin pigmentation is a complex process controlled by many different factors. Substance P (SP) regulates many biological functions, including melanogenesis and stress. Our previous study indicated that regulation of SP on melanocyte function was mediated by neurokinin 1 receptor (NK1 receptor). Substantial evidence has accumulated that psychological stress can be associated with skin pigmentation, so that the impact of 5-hydroxytryptamine (5-HT), one of the important factors participating in stress process, on melanogenesis has also been concerned. It has been reported that 5-HT induces melanin synthesis via 5-HT2A receptor. Furthermore, 5-HT2A receptor and NK1 receptor are G-protein coupled receptors (GPCRs) and both expressed on melanocyte, the present study was designed to investigate whether SP has influence on the adjustment function of 5-HT. Our data demonstrated that, SP inhibited 5-HT2A receptor expression to neutralize the pro-melanogenesis effect of 5-HT on B16F10 cells. The up-regulation of NK1 receptor expression was simultaneous with the down-regulation of 5-HT2A receptor treated by SP. This inhibition of 5-HT2A receptor expression by SP could be reversed by NK1 receptor antagonist Spantide I. Our studies indicated that SP could directly induce B16F10 cells apoptosis in vitro. 5-HT and 5-HT2A receptor agonist could mitigate this apoptotic effect of SP. It is the strong evidence of possible cross-talk between GPCRs and giving enlightenments when screening desirable drugs for target receptors. PMID:26872989

  7. {sup 124}I-BPA MicroPET images in C57BL/6 mouse bearing B16-F10 melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Chun Ki Jung [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of); Lee, Tae Sup; Woo, Kwang Sun; Chun, Kwon Soo; Cheon, Gi Jeon [Korea Institute Radiological and Medical Sciences, Seoul (Korea, Republic of)

    2007-07-01

    Boron Neutron Capture therapy(BNCT) uses a thermal or epithermal neutron source to bombard {sup 10}B atoms inside a cancer tissue and this produces short range alpha particles via a nuclear reaction that are able to kill tumor cells effectively. Preclinical and clinical trials have been conducted in the USA, Europe and Japan using {sup 10}B containing compounds such as BPA or BSH. This success will mainly depend on a differential uptake of {sup 10}B-BPA between a tumor and normal tissue. According to Solo way et al. (1998), a tumor-to-normal tissue uptake ratio (T/N) of 3:1 is desirable. However, it is difficult to directly measure {sup 10}B levels at the time of BNCT. Many researchers used radioactive analogs of {sup 10}B ([{sup 18}F]-FBPA) as a probe to analyze its kinetics in vivo using PET. However, the production of {sup 18}F-BPA is more difficult than that of {sup 124}I-BPA and its half-life is too short. In an earlier study, we obtained different individual images of C57BL/6 mice bearing melanoma on the thigh by a small animal PET scanner(microPET) using {sup 124}I-BPA instead of {sup 18}F-BPA on 1, 3 and 24 hr after an {sup 124}I-BPA injection and calculated the boron contents by the radioactivity in the tumor and the blood. In this study, the aim of our study is to obtain an image of a C57BL/6 mouse bearing melanoma by a small animal PET according to the time( 2 and 24hr) after an injection of a radiolabeled boron compound in order to obtain the changeable images by the injection time in the same individual.

  8. MicroPET image of radiolabeled boron compound {sup 124}I-BPA in C57BL/6 mouse bearing B16-F10 melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Chun, Ki-Jung; Kim, Woo-Jung [Korea Atomic Energy Research Institute, Taejon (Korea, Republic of); Kim, Mi-Sook; Chun, Kwon-Soo; Cheon, Gi-Jeong [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)

    2006-07-01

    Boron Neutron Capture therapy(BNCT) uses thermal or epithermal neutron source to bombard {sup 10}B atoms inside cancer tissue and this produces short-range alpha particles via a nuclear reaction that are able to kill tumor cells effectively. Preclinical and clinical trials have been conducted in the USA, Europe and Japan using {sup 10}B containing compounds such as BPA or BSH. This success will mainly depend on a differential uptake of {sup 10}B-BPA between tumor and normal tissue. According to Soloway et al. (1998), a tumor-to-normal tissue uptake ratio (T/N) of 3:1 is desirable. However, it is difficult to directly measure {sup 10}B levels at the time of BNCT. Many researchers used radioactive analogs of {sup 10}B ([{sup 18}F]-FBPA) as a probe to analyze its kinetics in vivo using PET. However, the production of {sup 18}FBPA is more difficult than that of {sup 124}I-BPA and its half life is too short. The aim of our study obtain to image C57BL/6 mice bearing melanoma on the thigh by a small animal PET scanner(microPET) using {sup 124}I-BPA instead of {sup 18}F-BPA and to calculate boron contents by the radioactivity in the tumor and the blood.

  9. Therapeutic T cells induce tumor-directed chemotaxis of innate immune cells through tumor-specific secretion of chemokines and stimulation of B16BL6 melanoma to secrete chemokines

    Directory of Open Access Journals (Sweden)

    Fox Bernard A

    2007-11-01

    Full Text Available Abstract Background The mechanisms by which tumor-specific T cells induce regression of established metastases are not fully characterized. In using the poorly immunogenic B16BL6-D5 (D5 melanoma model we reported that T cell-mediated tumor regression can occur independently of perforin, IFN-γ or the combination of both. Characterization of regressing pulmonary metastases identified macrophages as a major component of the cells infiltrating the tumor after adoptive transfer of effector T cells. This led us to hypothesize that macrophages played a central role in tumor regression following T-cell transfer. Here, we sought to determine the factors responsible for the infiltration of macrophages at the tumor site. Methods These studies used the poorly immunogenic D5 melanoma model. Tumor-specific effector T cells, generated from tumor vaccine-draining lymph nodes (TVDLN, were used for adoptive immunotherapy and in vitro analysis of chemokine expression. Cellular infiltrates into pulmonary metastases were determined by immunohistochemistry. Chemokine expression by the D5 melanoma following co-culture with T cells, IFN-γ or TNF-α was determined by RT-PCR and ELISA. Functional activity of chemokines was confirmed using a macrophage migration assay. T cell activation of macrophages to release nitric oxide (NO was determined using GRIES reagent. Results We observed that tumor-specific T cells with a type 1 cytokine profile also expressed message for and secreted RANTES, MIP-1α and MIP-1β following stimulation with specific tumor. Unexpectedly, D5 melanoma cells cultured with IFN-γ or TNF-α, two type 1 cytokines expressed by therapeutic T cells, secreted Keratinocyte Chemoattractant (KC, MCP-1, IP-10 and RANTES and expressed mRNA for MIG. The chemokines released by T cells and cytokine-stimulated tumor cells were functional and induced migration of the DJ2PM macrophage cell line. Additionally, tumor-specific stimulation of wt or perforin

  10. 10-Hydroxycamptothecin aerosol treatment inhibits lung metastases in B16F10 melanoma mice models%雾化吸入羟基喜树碱对小鼠B16黑色素瘤实验性肺转移的影响

    Institute of Scientific and Technical Information of China (English)

    张超; 胡巍; 方芸

    2011-01-01

    Objective: To estimate the effect of 10-hydroxycamptothecin (HCPT) to the melanoma lung metastasis mice models and the feasibility of aerosol delivery treatment for lung cancer therapy.Method: BI6FI0 melanoma lung metastasis mice models were made, and nodules number, inhibition rate, tumor area, mean nodules diameter and so on were investigated after the aerosol delivery treatment.Spleen index and thymus index were culculatad at the end of the experiments.The change of body weight, physiological state and the lung tumor tissue in pathological histology were inspeated.Result: The total number of tumor lesions, weight of lungs and the area of lung metastasis of aerosol treatment group had significant difference comparing with normal group and control group.Mean nodules diameter had no significant difference comparing with control group.The spleen index of aerosol treatment group was decreased and thymus index was significantly decreased comparing with normal group and control group.During the treatment there are no obvious changes in physiological state.The lung cancer tissue of aerosol delivery treatment group was recovered in pathological histology.Conclusion: The results suggested that aerosol delivery of HCPT demonstrated powerful antitumor activity and was useful for melanoma lung metastasis by aerosol delivery treatment.%目的:通过雾化10-羟基喜树碱(HCPT)用于治疗B16黑色素瘤肺转移癌小鼠,考察雾化吸入HCPT治疗肺癌的可行性以及疗效.方法:制作B16黑色素瘤肺转移癌小鼠模型,不同剂量雾化给药考察肺转移结节数、肺湿重、肺癌面积、平均直径等参数,计算脾脏指数和胸腺指数,并观察动物的生理生存状况,对肺癌组织进行病理组织学检查.结果:雾化给药组肺癌转移结节数、肺湿重、肺癌面积等参数明显少于模型组,直径相关参数未发生统计学差异;雾化给药组脾脏指数有所减小,胸腺指数较模型组和正常组有显著性差

  11. Natural CD8{sup +}25{sup +} regulatory T cell-secreted exosomes capable of suppressing cytotoxic T lymphocyte-mediated immunity against B16 melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Yufeng; Zhang, Xueshu; Zhao, Tuo; Li, Wei; Xiang, Jim, E-mail: jim.xiang@saskcancer.ca

    2013-08-16

    Highlights: •CD8{sup +}25{sup +} regulatory T cells secrete tolerogenic exosomes. •CD8{sup +}25{sup +} regulatory T cell-derived exosomes exhibit immunosuppressive effect. •CD8{sup +}25{sup +} regulatory T cell-derived exosomes inhibit antitumor immunity. -- Abstract: Natural CD4{sup +}25{sup +} and CD8{sup +}25{sup +} regulatory T (Tr) cells have been shown to inhibit autoimmune diseases. Immune cells secrete exosomes (EXOs), which are crucial for immune regulation. However, immunomodulatory effect of natural Tr cell-secreted EXOs is unknown. In this study, we purified natural CD8{sup +}25{sup +} Tr cells from C57BL/6 mouse naive CD8{sup +} T cells, and in vitro amplified them with CD3/CD28 beads. EXOs (EXO{sub Tr}) were purified from Tr cell’s culture supernatants by differential ultracentrifugation and analyzed by electron microscopy, Western blot and flow cytometry. Our data showed that EXO{sub Tr} had a “saucer” or round shape with 50–100 nm in diameter, contained EXO-associated markers LAMP-1 and CD9, and expressed natural Tr cell markers CD25 and GITR. To assess immunomodulatory effect, we i.v. immunized C57BL/6 mice with ovalbumin (OVA)-pulsed DCs (DC{sub OVA}) plus Tr cells or EXO{sub Tr}, and then assessed OVA-specific CD8{sup +} T cell responses using PE-H-2K{sup b}/OVA tetramer and FITC-anti-CD8 antibody staining by flow cytometry and antitumor immunity in immunized mice with challenge of OVA-expressing BL6–10{sub OVA} melanoma cells. We demonstrated that DC{sub OVA}-stimulated CD8{sup +} T cell responses and protective antitumor immunity significantly dropped from 2.52% to 1.08% and 1.81% (p < 0.05), and from 8/8 to 2/8 and 5/8 mice DC{sub OVA} (p < 0.05) in immunized mice with co-injection of Tr cells and EXO{sub Tr}, respectively. Our results indicate that natural CD8{sup +}25{sup +} Tr cell-released EXOs, alike CD8{sup +}25{sup +} Tr cells, can inhibit CD8{sup +} T cell responses and antitumor immunity. Therefore, EXOs derived from

  12. Natural CD8+25+ regulatory T cell-secreted exosomes capable of suppressing cytotoxic T lymphocyte-mediated immunity against B16 melanoma

    International Nuclear Information System (INIS)

    Highlights: •CD8+25+ regulatory T cells secrete tolerogenic exosomes. •CD8+25+ regulatory T cell-derived exosomes exhibit immunosuppressive effect. •CD8+25+ regulatory T cell-derived exosomes inhibit antitumor immunity. -- Abstract: Natural CD4+25+ and CD8+25+ regulatory T (Tr) cells have been shown to inhibit autoimmune diseases. Immune cells secrete exosomes (EXOs), which are crucial for immune regulation. However, immunomodulatory effect of natural Tr cell-secreted EXOs is unknown. In this study, we purified natural CD8+25+ Tr cells from C57BL/6 mouse naive CD8+ T cells, and in vitro amplified them with CD3/CD28 beads. EXOs (EXOTr) were purified from Tr cell’s culture supernatants by differential ultracentrifugation and analyzed by electron microscopy, Western blot and flow cytometry. Our data showed that EXOTr had a “saucer” or round shape with 50–100 nm in diameter, contained EXO-associated markers LAMP-1 and CD9, and expressed natural Tr cell markers CD25 and GITR. To assess immunomodulatory effect, we i.v. immunized C57BL/6 mice with ovalbumin (OVA)-pulsed DCs (DCOVA) plus Tr cells or EXOTr, and then assessed OVA-specific CD8+ T cell responses using PE-H-2Kb/OVA tetramer and FITC-anti-CD8 antibody staining by flow cytometry and antitumor immunity in immunized mice with challenge of OVA-expressing BL6–10OVA melanoma cells. We demonstrated that DCOVA-stimulated CD8+ T cell responses and protective antitumor immunity significantly dropped from 2.52% to 1.08% and 1.81% (p OVA (p Tr, respectively. Our results indicate that natural CD8+25+ Tr cell-released EXOs, alike CD8+25+ Tr cells, can inhibit CD8+ T cell responses and antitumor immunity. Therefore, EXOs derived from natural CD4+25+ and CD8+25+ Tr cells may become an alternative for immunotherapy of autoimmune diseases

  13. In vitro and in vivo studies of the antineoplastic activity of copper (II) compounds against human leukemia THP-1 and murine melanoma B16-F10 cell lines.

    Science.gov (United States)

    Borges, Layla J H; Bull, Érika S; Fernandes, Christiane; Horn, Adolfo; Azeredo, Nathalia F; Resende, Jackson A L C; Freitas, William R; Carvalho, Eulógio C Q; Lemos, Luciana S; Jerdy, Hassan; Kanashiro, Milton M

    2016-11-10

    We investigated the antineoplastic activities of a previously reported copper (II) coordination compound, [Cu(BMPA)Cl2]CH3OH (1), and a new compound, [Cu(HBPA)Cl2]H2O (2), where BMPA is bis(pyridin-2-ylmethyl)amine and HBPA is (2-hydroxybenzyl)(2-pyridylmethyl)amine, using various cellular models of human leukemia (THP-1, U937, HL60, Molt-4, JURKAT) and human colon cancer (COLO 205), as well as a murine highly metastatic melanoma (B16-F10) cell line. Compound (2) was characterized using several physical and chemical techniques, including X-ray diffraction studies. The IC50 values of the copper coordination complexes in the human leukemia cell lines ranged from 87.63 ± 1.02 to ≥400 μM at high cell concentrations and from 19.17 ± 1.06 to 97.67 ± 1.23 μM at low cell concentrations. Both compounds induced cell death, which was determined by cell cycle analyses and phosphatidylserine exposure studies. THP-1 cells released cytochrome c to the cytoplasm 12 h after treatment with 400 μM of compound (2). To evaluate the apoptosis pathway induced by compound (2), we measured the activities of initiator caspases 8 and 9 and executioner caspases 3 and 6. The results were suggestive of the activation of both intrinsic and extrinsic apoptosis pathways. To investigate the activities of the compounds in vivo, we selected two sensitive cell lines from leukemia (THP-1) and solid tumor (B16-F10) lineages. BALB/c nude bearing THP-1 tumors treated with 12 mg·kg(-1) of compound (2) showed a 92.4% inhibition of tumor growth compared with the control group.

  14. Glucocorticoid receptor knockdown decreases the antioxidant protection of B16 melanoma cells: an endocrine system-related mechanism that compromises metastatic cell resistance to vascular endothelium-induced tumor cytotoxicity.

    Science.gov (United States)

    Obrador, Elena; Valles, Soraya L; Benlloch, María; Sirerol, J Antoni; Pellicer, José A; Alcácer, Javier; Coronado, Javier Alcácer-F; Estrela, José M

    2014-01-01

    We previously reported an interorgan system in which stress-related hormones (corticosterone and noradrenaline), interleukin-6, and glutathione (GSH) coordinately regulate metastatic growth of highly aggressive B16-F10 melanoma cells. Corticosterone, at levels measured in tumor-bearing mice, also induces apoptotic cell death in metastatic cells with low GSH content. In the present study we explored the potential role of glucocorticoids in the regulation of metastatic cell death/survival during the early stages of organ invasion. Glucocorticoid receptor (GCR) knockdown decreased the expression and activity of γ-glutamylcysteine synthetase (γ-GCS), the rate-limiting step in GSH synthesis, in metastatic cells in vivo independent of the tumor location (liver, lung, or subcutaneous). The decrease in γ-GCS activity was associated with lower intracellular GSH levels. Nrf2- and p53-dependent down-regulation of γ-GCS was associated with a decrease in the activities of superoxide dismutase 1 and 2, catalase, glutathione peroxidase, and glutathione reductase, but not of the O2--generating NADPH oxidase. The GCR knockdown-induced decrease in antioxidant protection caused a drastic decrease in the survival of metastatic cells during their interaction with endothelial cells, both in vitro and in vivo; only 10% of cancer cells attached to the endothelium survived compared to 90% survival observed in the controls. This very low rate of metastatic cell survival was partially increased (up to 52%) in vivo by inoculating B16-F10 cells preloaded with GSH ester, which enters the cell and delivers free GSH. Taken together, our results indicate that glucocorticoid signaling influences the survival of metastatic cells during their interaction with the vascular endothelium.

  15. New Sesquiterpene Lactone Dimer, Uvedafolin, Extracted from Eight Yacon Leaf Varieties (Smallanthus sonchifolius): Cytotoxicity in HeLa, HL-60, and Murine B16-F10 Melanoma Cell Lines.

    Science.gov (United States)

    Kitai, Yurika; Hayashi, Kana; Otsuka, Moe; Nishiwaki, Hisashi; Senoo, Tatsuya; Ishii, Tomohiko; Sakane, Genta; Sugiura, Makoto; Tamura, Hirotoshi

    2015-12-23

    Uvedafolin, 1, a new sesquiterpene lactone dimer, was isolated from the leaves of Smallanthus sonchifolius with five related compounds, 2-6, and their cytotoxicity was assessed against three tumor cell lines (HeLa, HL-60, B16-F10 melanoma). The stereostructure of 1 was newly elucidated by ESI-TOF-MS, 1D/2D NMR, and single-crystal X-ray diffraction. Dimers 1 and 2 had the most effective IC50 values, 0.2-1.9 μM, against the three tumor cell lines when compared with monomers 3-6 (IC50 values 0.7-9.9 μM) and etoposide (IC50 values 0.8-114 μM). The ester linkages of two sets of monomers, uvedalin, 5, and sonchifolin, 6, for 1, and enhydrin, 4, and sonchifolin, 6, for 2, as well as the acetyl group at the C-9 position, were essential for the high cytotoxicity. Dimers 1 and 2 would have potential as anticancer agents. PMID:26576855

  16. Syntaxin 7 complexes with mouse Vps10p tail interactor 1b, syntaxin 6, vesicle-associated membrane protein (VAMP)8, and VAMP7 in b16 melanoma cells.

    Science.gov (United States)

    Wade, N; Bryant, N J; Connolly, L M; Simpson, R J; Luzio, J P; Piper, R C; James, D E

    2001-06-01

    Syntaxin 7 is a mammalian target soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) involved in membrane transport between late endosomes and lysosomes. The aim of the present study was to use immunoaffinity techniques to identify proteins that interact with Syntaxin 7. We reasoned that this would be facilitated by the use of cells producing high levels of Syntaxin 7. Screening of a large number of tissues and cell lines revealed that Syntaxin 7 is expressed at very high levels in B16 melanoma cells. Moreover, the expression of Syntaxin 7 increased in these cells as they underwent melanogenesis. From a large scale Syntaxin 7 immunoprecipitation, we have identified six polypeptides using a combination of electrospray mass spectrometry and immunoblotting. These polypeptides corresponded to Syntaxin 7, Syntaxin 6, mouse Vps10p tail interactor 1b (mVti1b), alpha-synaptosome-associated protein (SNAP), vesicle-associated membrane protein (VAMP)8, VAMP7, and the protein phosphatase 1M regulatory subunit. We also observed partial colocalization between Syntaxin 6 and Syntaxin 7, between Syntaxin 6 and mVti1b, but not between Syntaxin 6 and the early endosomal t-SNARE Syntaxin 13. Based on these and data reported previously, we propose that Syntaxin 7/mVti1b/Syntaxin 6 may form discrete SNARE complexes with either VAMP7 or VAMP8 to regulate fusion events within the late endosomal pathway and that these events may play a critical role in melanogenesis.

  17. Effects of Supernatant of CLEC2B Gene Overexpression in Jurkat Cells on the B16 Melanoma cell%CLEC2B基因过表达的Jurkat细胞培养上清液对黑素瘤细胞B16的影响

    Institute of Scientific and Technical Information of China (English)

    张峻岭; 徐士福; 柳君如; 程琳; ZHOU Youwen

    2012-01-01

    目的 将CLEC2B基因过表达的Jurkat细胞培养上清液作用于黑素瘤细胞,观察其上清液对黑素瘤细胞增殖、酪氨酸酶活性、黑素合成的影响,探讨CLEC2B基因在白癜风发病中的作用.方法 培养人淋巴瘤细胞Jurkat细胞和小鼠黑素瘤细胞B16,采用脂质体介导的方法瞬时转染CLEC2B重组质粒入Jurkat细胞,半定量RT-PCR法鉴定CLEC2B基因过表达;将其细胞培养上清液作用于黑素瘤细胞48h后,MTT法检测黑素瘤细胞的增殖情况,多巴氧化法检测酪氨酸酶活性,氢氧化钠裂解法检测黑素含量.结果 CLEC2B基因过表达的Jurkat细胞培养上清液作用后的黑素瘤细胞增殖比空载体组、正常对照组降低,差异有统计学意义(均P<0.05),CLEC2B过表达组黑素含量比空载体组、正常对照组减少,差异有统计学意义(均P<0.05),CLEC2B过表达组、空载体组、正常对照组之间的酪氨酸酶活性比较差异无统计学意义(均P>0.05),空载体组与正常对照组比较差异均无统计学意义(均P>0.05).结论 CLEC2B基因过表达的Jurkat细胞培养上清液对黑素细胞的增殖和黑素合成有一定的抑制作用,对酪氨酸酶活性影响不明显.提示CLEC2B可能通过调节淋巴细胞功能间接影响黑素细胞增殖和黑素合成,从而参与白癜风发病.%Objective The melanoma cells were treated with culture supernatant of CLEC2B gene overexpression in Jurkat cells, to observe the impact of melanoma proliferation, the activity of tyrosinase and melanin synthesis, for searching CLEC2B gene participated in the vitiligo. Methods In this study, Jurkat cells were transfected with recombinant plasmid pGCMV/EGFP/Neo-CLEC2B by DMRIE-C Reagent. RT-PCR identified CLEC2B gene overexpression. After 48 h transfection, the melanoma cells were treated with their culture supernatant for 48 h, then the proliferation of melanoma cells were detected by MTT, the activity of tyrosinase was detected by

  18. Myeloid Cells' Evasion of Melanoma Immunity

    Science.gov (United States)

    Wang, Jun; Chen, Lieping

    2015-01-01

    An immune-suppressive role of myeloid-derived suppressor cells (MDSCs) in melanoma has long been speculated, whereas molecular mechanisms underlying this role are not well understood. Here, Chung and colleagues show that dendritic cell-associated, heparan sulfate proteoglycans-dependent integrin ligand (DC-HIL), a cell surface immune-modulatory molecule, is highly expressed on tumor-associated MDSCs. Genetic ablation or antibody blockade of DC-HIL delays the growth of transplantable B16 melanoma in syngeneic mice, which is accompanied by enhanced antitumor T-cell activities. These findings support a role for DC-HIL in immune evasion within the melanoma microenvironment. PMID:25318429

  19. Chlorpromazine distribution in hamsters and mice bearing transplantable melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Fairchild, R.G. (Brookhaven National Lab., Upton, NY); Greenberg, D.; Watts, K.P.; Packer, S.; Atkins, H.L.; Som, P.; Hannon, S.J.; Brill, A.B.; Fand, I.; McNally, W.P.

    1982-02-01

    Chlorpromazine (CPZ) distribution was measured in tissues of Syrian golden hamsters bearing Greene melanoma and in BALB/c mice bearing Harding-Passey melanoma. Distribution was evaluated as a function of time (0.5 to 14 days) and as a function of single and multiple doses (up to five) of from 5 to 50 mg CPZ per kg body weight. Routes of administration (i.p., i.v., p.o.) were compared. The physiological behavior of CPZ is of interest as it is used extensively as a tranquilizing drug (Thorazine). Further, since CPZ binds to the pigment melanin, the possibility exists of using CPZ to transport diagnostic or therapeutic agents to melanoma. It was found that, at 2 days postinjection, tumor/tissue concentration ratios exceeded 10 for metabolizing organs, such as liver, and 100 for background tissues, such as blood and muscle. Absolute concentrations of CPZ in tumor exceeding 100 ..mu..g CPZ per g tumor were obtained with both single and multiple doses. This selective high concentration in tumor would make CPZ an ideal vehicle for the transport of boron to tumor for use in neutron capture therapy via the /sup 10/B(n,..cap alpha..)/sup 7/Li reaction.

  20. Melanoma

    Science.gov (United States)

    ... may be melanoma is the ABCDE checklist: A – Asymmetry: One half of the mole does not look ... 276. PMID: 16050450. Last Updated: 12 Feb 2009 Information for other ages: Table of Contents: Overview Who's ...

  1. Overview on non-melanoma skin cancers in solid organ transplant recipients.

    Science.gov (United States)

    Forchetti, G; Suppa, M; Del Marmol, V

    2014-08-01

    The risk of non-melanoma skin cancer (NMSC) is significantly increased in solid organ transplant recipients (SOTRs) due to the long-term immunosuppressive treatment. NMSCs can be more aggressive in SOTRs than in the general population, resulting in significantly higher morbidity and mortality. In contrast to the immunocompetent population, skin cancers in SOTRs are dominated by squamous cell carcinoma, followed by basal cell carcinoma. Life-long radiation exposure, male sex, fair skin, history of prior NMSC, genetic factors, age at transplant along with duration and extent of the immunosuppression therapy have been identified as risk factors for NMSC in SOTRs. Photo-protection, skin self-examination, early diagnosis and treatment of skin lesions, reduction of immunotherapy, switch to mammalian target-of-rapamycin inhibitors and chemoprevention with oral retinoids are effective measures for the reduction of the incidence of NMSC in such patients. PMID:25068224

  2. Growth of transplantable melanoma and leukaemia and prevention of virus-induced leukaemia in long lived radiation chimeras constructed with unmanipulated bone marrow

    International Nuclear Information System (INIS)

    Haemopoietic radiation chimeras across the H-2 barrier (BALB/c → C57B1/6; H-2sup(d) → H-2sup(b) chimeras and vice versa) have been studied for their capacity to suppress the growth, or to reject, transplantable B16 melanotic melanoma and radiation leukaemia virus-induced, transplantable leukaemia. Also, radiation leukaemia virus (RadLV) obtained from the thymus of leukaemic C57B1/6 mice was injected i.p. into established chimeras (H-2sup(d) → H-2sup(b)). As expected, long lived, graft versus host disease free allogeneic chimeras constructed with intact bone marrow were unable to reject the tumours both when recipients were BALB/c → C57B1/6 or C57B1/6 → BALB/c chimeras. However, inoculation of a large number of immunocompetent cells from normal BALB/c mice into BALB/c → C57B1/6 chimeras failed to promote a rejection of the tumours. On the contrary, the same amount of syngeneic (BALB/c) immunocompetent cells prevented growth of melanoma when transferred into athymic nude BALB/c mice, while the tumour grew unimpaired in untreated athymic nude BALB/c mice. The same type of H-2sup(d) → H-2sup(b) chimeras displayed complete resistance to inoculation of leukaemogenic H-2sup(b) restricted RadLV while all H-2sup(b) → H-2sup(b), syngeneically reconstituted mice developed disseminated leukaemia. (author)

  3. 复方木尼孜其颗粒对小鼠B16黑素瘤细胞酪氨酸酶活性及黑素合成的影响%Effects of Fufang Muniziqi Particle on Melanin Synthesis and Tyrosinase Activity in Mouse B16 Melanoma Cells

    Institute of Scientific and Technical Information of China (English)

    纪超; 毕志刚

    2014-01-01

    目的 研究复方木尼孜其颗粒对小鼠B16黑素瘤细胞酪氨酸酶活性及黑素合成的影响.方法 培养小鼠B16黑素瘤细胞,采用MTT法测定不同浓度复方木尼孜其颗粒对细胞增殖的影响,采用L-DOPA氧化法测定其对细胞酪氨酸酶活性的影响,采用NaOH裂解法检测细胞黑素含量.结果 复方木尼孜其颗粒高剂量组能明显抑制小鼠B16黑素瘤细胞黑素的合成,但是对酪氨酸酶活性无影响.结论 复方木尼孜其颗粒能抑制小鼠B16黑素瘤细胞黑素的合成,在治疗色素性皮肤病中有一定前景.

  4. 杏仁油对黑色素瘤细胞B16细胞增殖及黑素合成的影响%Effects of Almond Kernel Oil on Cell Proliferation and Melanin Synthesis of Cultured B16 Murine Melanoma Cell

    Institute of Scientific and Technical Information of China (English)

    任燕冬; 宋宏杉; 陈巧云; 张宁

    2011-01-01

    目的 探讨杏仁油对小鼠黑色素瘤细胞B16增殖及黑素合成的影响.方法 采用MTT法测定杏仁油对小鼠黑色素瘤细胞B16增殖的影响,L-dopa氧化法测定酪氨酸酶活性,NaOH裂解法测定黑素合成.结果 杏仁油浓度在0.1~100μg/mL时对小鼠黑色素瘤细胞B16增殖无影响(P>0.05),对酪氨酸酶活性和黑素含量均呈浓度依赖性抑制,均具有显著性抑制作用(P<0.05).结论 杏仁油对黑色素瘤细胞B16酪氨酸酶活性和黑素生成有较强的剂量相关性抑制作用.

  5. 蟛蜞菊乙醇提取物促进黑色素生成及其机理的初步研究%The effect and mechanism of ethanol extract from Wedelia chinensis on melanogenesis of B16 melanoma cells

    Institute of Scientific and Technical Information of China (English)

    梅寒芳; 林密; 朱家勇

    2012-01-01

    Objective To study the effect and potential mechanism of ethanol extracts from Wedelia chinensis (EEW) on melanogenesis of B16 murine melanoma cells. Methods B16 cells were incubated with different concentrations of EEW for 72 h,and then cell viability was measured by MTT method, tyrosinase activity by L-dopa oxidative reaction, and melanogenesis by NaOH method, respectively. The mRNA expression of tyrosinase and microphthalmia associated transcription factor (MITF) were detected by RT-PCR. Results In the range of experimental dose, EEW slightly increased cell viability. The activity of tyrosinase and production of melanin were significantly increased in a dose-dependent manner. The increased gene expression of tyrosinase and MITF were also observed in a dose-dependent manner. Conclusion EEW can directly stimulate melanogenesis in B16 melanoma cells, which may be mediated by promoting cell proliferation and increasing expression of tyrosinase and MITF.%目的 观察中药蟛蜞菊乙醇提取物促进黑色素生成的作用,并初步探讨其作用机理.方法 MTT法、L-Dopa氧化法、NaOH裂解法探讨不同浓度蟛蜞菊乙醇提取物作用于小鼠恶性黑色素瘤细胞(B16)72 h后,对细胞增殖、酪氨酸酶活性及黑色素含量的影响.RT-PCR检测药物作用前后酪氨酸酶、小眼相关转录因子(MITF)等基因表达的影响.结果 与对照组比较,蟛蜞菊乙醇提取物能促进B16细胞的增殖,促进酪氨酸酶活性增加和黑色素生成增多;可剂量依赖性上调酪氨酸酶和MITF的mRNA表达.结论 蟛蜞菊乙醇提取物可促进B16细胞黑色素生成,其机制可能是通过促进B16细胞增殖、增强酪氨酸酶和MITF的基因表达来实现.

  6. Human melanoma cells transplanted into zebrafish proliferate, migrate, produce melanin, form masses and stimulate angiogenesis in zebrafish.

    Science.gov (United States)

    Haldi, Maryann; Ton, Christopher; Seng, Wen Lin; McGrath, Patricia

    2006-01-01

    In this research, we optimized parameters for xenotransplanting WM-266-4, a metastatic melanoma cell line, including zebrafish site and stage for transplantation, number of cells, injection method, and zebrafish incubation temperature. Melanoma cells proliferated, migrated and formed masses in vivo. We transplanted two additional cancer cell lines, SW620, a colorectal cancer cell line, and FG CAS/Crk, a pancreatic cancer cell line and these human cancers also formed masses in zebrafish. We also transplanted CCD-1092Sk, a human fibroblast cell line established from normal foreskin and this cell line migrated, but did not proliferate or form masses. We quantified the number of proliferating melanoma and normal skin fibroblasts by dissociating xenotransplant zebrafish, dispensing an aliquot of CM-DiI labeled human cells from each zebrafish onto a hemocytometer slide and then visually counting the number of fluorescently labeled cancer cells. Since zebrafish are transparent until approximately 30 dpf, the interaction of labeled melanoma cells and zebrafish endothelial cells (EC) can be visualized by whole-mount immunochemical staining. After staining with Phy-V, a mouse anti-zebrafish monoclonal antibody (mAb) that specifically labels activated EC and angioblasts, using immunohistology and 2-photon microscopy, we observed activated zebrafish EC embedded in human melanoma cell masses. The zebrafish model offers a rapid efficient approach for assessing human cancer cells at various stages of tumorigenesis. PMID:17051341

  7. An ethical dilemma: malignant melanoma in a 51-year-old patient awaiting simultaneous kidney and pancreas transplantation for type 1 diabetes.

    Science.gov (United States)

    Kirby, L C; Banerjee, A; Augustine, T; Douglas, J F

    2016-07-01

    Malignant melanoma is a high-risk skin cancer that, in potential transplant recipients, is considered a substantial contraindication to solid organ transplantation due to significant risk of recurrence with immunosuppression. Current guidelines stipulate waiting between 3 and 10 years after melanoma diagnosis. However, in young patients with end-stage organ failure and malignant melanoma, complex ethical and moral issues arise. Assessment of the true risk associated with transplantation in these patients is difficult due to lack of prospective data, but an autonomous patient can make a decision that clinicians may perceive to be high risk. The national and worldwide shortage of available organs also has to be incorporated into the decision to maximize the net benefit and minimize the risk of graft failure and mortality. The incidence of malignant melanoma worldwide is increasing faster than that of any other cancer and continues to pose ethically challenging decisions for transplant specialists evaluating recipients for solid organ transplantation. PMID:27484276

  8. 黑芝麻提取物促B16黑素瘤细胞黑素合成及其机制的研究%The Effect of Water Extract from Semen Sesami Nigrum on Melanogenesis of B16 Melanoma Cells

    Institute of Scientific and Technical Information of China (English)

    姜泽群; 徐继敏; 吴琼; 何光源

    2009-01-01

    目的 研究黑芝麻的水提取物对B16黑素瘤细胞系酪氨酸酶活性、黑素合成及相关基因和蛋白表达的影响,探讨黑芝麻促进黑色素合成的可能机制.方法 选择3种浓度(0.5,1,2 mg/ml)的黑芝麻水提物作用于B16黑素瘤细胞72 h,观察其对黑素细胞的增殖、酪氨酸酶活性和黑素含量的影响.免疫印迹法和半定量逆转录聚合酶链式反应(RT-PCR)分别检测药物作用48 h前后酪氨酸酶(Tyrosinase)、小眼相关转录因子(microphthalmia associated transcription factor,MITF),酪氨酸酶相关蛋白酶1(TRPl)和酪氨酸酶相关蛋白酶2(TRP2)蛋白和mRNA表达水平的变化.结果 3种浓度的黑芝麻水提物可轻微促进B16细胞的增殖(P> 0.05);试验浓度范围内可明显促进B16细胞黑素合成和酪氨酸酶活性增加( P 0.05).结论 黑芝麻水提物在体外能直接刺激B16黑素瘤细胞黑色素的合成,上述变化可能通过促进酪氨酸酶和MITF的基因转录和蛋白表达作用来完成.

  9. 鱿鱼皮胶原蛋白多肽对B16黑素瘤细胞黑素合成的影响%Effects of collagen polypeptides from squid (Dosidicus gigas)skin on melanogenesis in B16 melanoma cells

    Institute of Scientific and Technical Information of China (English)

    王静凤; 王奕; 崔凤霞; 李八方; 薛长湖

    2007-01-01

    目的 研究不同分子量鱿鱼皮胶原蛋白多肽SP1(Mr>10 000u)、SP2(6 000u<Mr<10 000u)、SP3(2 000u<Mr<6 000u)对小鼠B16黑素瘤细胞黑素含量、酪氨酸酶活性及酪氨酸酶基因表达的影响.方法 采用MTT法测定细胞增殖活性,NaOH裂解法测定黑素含量,多巴氧化法测定酪氨酸酶活性,逆转录-聚合酶链反应(RT-PCR)测定酪氨酸酶mRNA表达.结果 不同分子量鱿鱼皮胶原蛋白多肽均能明显降低B16黑素瘤细胞黑素含量(P<0.05,P<0.01),抑制酪氨酸酶活性(P<0.01),并下调酪氨酸酶mRNA表达(P<0.05,P<0.01),且剂量效应关系明显.结论 不同分子量鱿鱼皮胶原蛋白多肽均具有明显抑制B16黑素瘤细胞黑素合成的作用,其中SP2的抑制效果较SP1、SP3明显(P<0.05).

  10. 熊果苷和甘草黄酮对B16黑素瘤细胞株黑素合成的影响%The Comparative Study of Arbutine and Glabridin on Regulation of Melanogenesis in B16 Murine Melanoma Cells

    Institute of Scientific and Technical Information of China (English)

    吴品茹; 徐慧; 陈向东; 沈征宇; 刘健航

    2008-01-01

    目的 比较观察甘草黄酮、熊果苷对体外培养的B16鼠黑素瘤细胞黑素合成的影响.方法 选择系列浓度梯度的药物作用于B16黑素瘤细胞株,测定细胞酪氨酸酶活性、黑素含量和细胞增殖率变化.结果 在实验浓度时,甘草黄酮具有浓度依赖性黑素合成抑制作用,与熊果苷比较,差异有统计学意义(P<0.05).结论 两种化合物均显示有抑制黑素生成的作用,存在一定细胞毒性.

  11. 蛇毒半胱氨酸蛋白酶抑制剂的酵母表达及其对B16细胞体外侵袭的抑制作用%Expression of Recombinant Snake Venom Cystatin in Yeast Pichia pastoris and Its Effects on B16F1 Melanoma Invasion in vitro

    Institute of Scientific and Technical Information of China (English)

    万榕; 宋军; 郑海音; 张晓艳; 林旭; 林建银

    2007-01-01

    目的:建立毕赤酵母表达质粒pPICZαA-cystatin,转化酵母细胞生产重组蛋白,探讨蛇毒半胱氨酸蛋白酶抑制剂(sv-cystatin)对肿瘤侵袭的生物学作用.方法:利用PCR扩增技术从pUC18/sv-cystatin质粒中扩增sv-cystatin cDNA并克隆至酵母表达载体pPICZαA上,构建重组质粒pPICZαA-cystatin电激转化Pichia pastori酵母细胞GS115,经1%甲醇诱导获得稳定表达的重组蛋白,改良Boyden小室分析重组sv-cystatin蛋白处理对B16F1细胞体外侵袭力的影响.结果:SDS-PAGE检测和Western blot分析显示分泌表达的sv-cystatin重组蛋白相对分子量约为14kDa,摇瓶发酵每升发酵培养上清可获得16 mg的重组蛋白,经亲和层析纯化获得的sv-cystatin重组蛋白具有抑制木瓜蛋白酶的活性.改良Boyden小室实验结果显示:经0.5mg/ml浓度的重组蛋白处理的B16F1细胞穿过Matrigel的细胞数明显低于对照组(52.60±4.58,106±5.9,P<0.01) ,抑制率为50%.结论:成功实现sv-cystatin的酵母表达,初步证明sv-cystatin重组蛋白可抑制小鼠B16F1细胞体外侵袭作用.

  12. Recommendations for Solid Organ Transplantation for Transplant Candidates With a Pretransplant Diagnosis of Cutaneous Squamous Cell Carcinoma, Merkel Cell Carcinoma and Melanoma: A Consensus Opinion From the International Transplant Skin Cancer Collaborative (ITSCC).

    Science.gov (United States)

    Zwald, F; Leitenberger, J; Zeitouni, N; Soon, S; Brewer, J; Arron, S; Bordeaux, J; Chung, C; Abdelmalek, M; Billingsley, E; Vidimos, A; Stasko, T

    2016-02-01

    Advancements in solid organ transplantation successfully extend the lives of thousands of patients annually. The tenet of organ stewardship aims to prevent the futile expenditure of scarce donor organs in patient populations with high mortality risk, to the detriment of potential recipients with greater predicted life expectancy. The development of skin cancer posttransplantation portends tremendous morbidity, adversely affecting quality of life for many transplant recipients. This special article, provided by of members of the International Transplant Skin Cancer Collaborative (ITSCC), will provide the transplant professional with a consensus opinion and recommendations as to an appropriate wait period pretransplantation for transplant candidates with a history of either cutaneous squamous cell carcinoma, malignant melanoma, or Merkel cell carcinoma. PMID:26820755

  13. Role of radiotherapy in the management of organ transplant recipients diagnosed with non-melanoma skin cancers

    International Nuclear Information System (INIS)

    Organ transplantation has had a major effect on the lives of thousands of patients worldwide. In Australia and New Zealand, over 13 000 patients have become organ transplant recipients (OTR). Following transplantation, patients require lifelong immunosuppression to prevent organ rejection. The loss of immune surveillance results in OTR experiencing a higher incidence of infection and malignancy in comparison with the general (immunocompetent) population. Non-melanoma skin cancer (NMSC) is the most common malignancy worldwide, arising most often on the sun-exposed head and neck. Organ transplant recipients experience a higher incidence of NMSC when compared with the general population and a higher incidence of squamous cell carcinoma compared with basal cell carcinoma. Organ transplant recipients also develop NMSC at a younger age and experience multiple new NMSC. Australians experience the highest incidence of NMSC in the world with a consequence that NMSC arising in OTR can lead to significant morbidity and even mortality. Radiation oncologists treating patients with skin cancer will almost certainly make recommendations in the setting of NMSC arising in OTR. The aim of this article is to discuss the role of radiotherapy in the management of OTR diagnosed with NMSC. The emphasis will be on the treatment of patients with a high-risk NMSC (e.g. squamous cell carcinoma, Merkel cell carcinoma, unfavourable basal cell carcinoma) because this reflects the most common clinical scenario in which a recommendation of radiotherapy, usually adjuvant, may be considered

  14. Antitumoral, antioxidant, and antimelanogenesis potencies of Hawthorn, a potential natural agent in the treatment of melanoma.

    Science.gov (United States)

    Mustapha, Nadia; Mokdad-Bzéouich, Imèn; Maatouk, Mouna; Ghedira, Kamel; Hennebelle, Thierry; Chekir-Ghedira, Leila

    2016-06-01

    The lack of an efficient agent that does not have the disadvantage of low activity (kojic acid), high cytotoxicity, and mutagenicity (hydroquinone), poor skin penetration (arbutin), or low stability in formulation (glabridin) led us to continue our research on new antipigmentation/skin-lightening agents. Therefore, research of natural products that can modulate the metabolism of pigmentation is of great interest. Otherwise, malignant melanoma is one of the most aggressive forms of skin cancer, with high metastatic potential, and currently, there is no effective chemotherapy against invasive melanoma. Therefore, it is necessary to develop new drugs with potent activity and weak side effects against melanoma. The in-vitro anticancer effect of hawthorn was analyzed against B16F10 melanoma cells using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The effect of isolated compounds from hawthorn on melanogenesis in B16F10 melanoma cells was investigated by measuring the amounts of melanin and tyrosinase spectrophotometrically at 475 nm. Balb/c mice models inoculated with B16F10 mouse tumor cells were used to evaluate the in-vivo antitumoral potential of hawthorn by assessing its effect on the growth of transplanted tumors. The antioxidant potential of tested samples was evaluated in B16F10 and primary human keratinocyte cells using a cellular antioxidant activity assay. Hawthorn tested samples inhibited effectively the growth of melanoma cells in vitro. Furthermore, it appears that tested samples from hawthorn reduced melanogenesis by inhibiting the tyrosinase activity of B16F10 cells in a dose-dependent manner. In-vivo studies showed that hawthorn total oligomer flavonoids extract treatment at a dose of 150 mg/kg body weight for 21 days in implanted tumor mice resulted in significant inhibition of the tumor growth volume and weight. In addition, tested samples showed significant cellular antioxidant capacity against the reactive oxygen species

  15. Lentivirus-mediated bifunctional cell labeling for in vivo melanoma study.

    Science.gov (United States)

    Day, Chi-Ping; Carter, John; Bonomi, Carrie; Esposito, Dominic; Crise, Bruce; Ortiz-Conde, Betty; Hollingshead, Melinda; Merlino, Glenn

    2009-06-01

    Lentiviral vectors (LVs) are capable of labeling a broad spectrum of cell types, achieving stable expression of transgenes. However, for in vivo studies, the duration of marker gene expression has been highly variable. We have developed a series of LVs harboring different promoters for expressing reporter gene in mouse cells. Long-term culture and colony formation of several LV-labeled mouse melanoma cells showed that promoters derived from mammalian house-keeping genes, especially those encoding RNA polymerase II (Pol2) and ferritin (FerH), provided the highest consistency for reporter expression. For in vivo studies, primary B16BL6 mouse melanoma were infected with LVs whose luciferase-green fluorescence protein fusion gene (Luc/GFP) was driven by either Pol2 or FerH promoters. When transplanted into syngeneic C57BL/6 mice, Luc/GFP-labeled B16BL6 mouse melanoma cells can be monitored by bioluminescence imaging in vivo, and GFP-positive cells can be isolated from the tumors by fluorescence-activated cell sorter. Pol2-Luc/GFP labeling, while lower in activity, was more sustainable than FerH-Luc/GFP labeling in B16BL6 over consecutive passages into mice. We conclude that Pol-2-Luc/GFP labeling allows long-term in vivo monitoring and tumor cell isolation in immunocompetent mouse melanoma models. PMID:19175523

  16. Melanoma of the eye

    Science.gov (United States)

    ... modified July 9, 2015. www.cancer.gov/types/eye/hp/intraocular-melanoma-treatment-pdq . Accessed October 7, 2015. Read ... by: Yi-Bin Chen, MD, Leukemia/Bone Marrow Transplant Program, Massachusetts General Hospital, Boston, MA. Also reviewed ...

  17. Radioiodinated N-(alkylaminoalkyl)-substituted 4-methoxy-, 4-hydroxy-, and 4-aminobenzamides: Biological investigations for the improvement of melanoma-imaging agents

    International Nuclear Information System (INIS)

    The synthesis and radiolabeling of 27 new N-(alkylaminoalkyl)-4-methoxy-, 4-hydroxy-, and -4-aminobenzamides are described and evaluated in C57Bl/6 mice with subcutaneously transplanted B16 melanoma in order to screen the optimal chemical structure for melanoma scintigraphy. Using Tl(TFA)3 for 131I- labeling, a series of radioiodinated 4-methoxy benzamide derivatives proved to exhibit superior melanoma uptake with outstanding melanoma/non-target-tissue ratios. From the benzamide derivatives tested, N-(2-(1'-piperidinyl)ethyl)-3-[131I]iodo-4-methoxybenzamide (1) and N-(2-diethylaminoethyl)-3-[131I]iodo-4-methoxybenzamide (2) demonstrated best results. The introduction of 4-hydroxy and 4-amino groups led to less favourable benzamides. While the former compounds showed little melanoma uptake, the latter revealed unfavourable melanoma/non-target-tissue ratios. Additionally, it could be shown that an amino group was inevitably necessary for melanoma uptake, and that dialkylation of the amide nitrogen and replacing CONH by CH2NH revealed less advantageous results

  18. Enucleaton of the right eye due to large choroidal melanoma with simultaneous penetrating cornea transplantation from OD to OS (Case report.

    Directory of Open Access Journals (Sweden)

    E. A. Korchuganova

    2013-01-01

    Full Text Available There is presentation case report of 75‑year old woman with choroidal melanoma (T3N0M0 on the right eye and failed graft on the left pseudophakic eye with far advanced glaucoma and ARMD. No treatment was given to the leading eye with VA 0,2. VA of OS = 1 / ∞ pr.l.certa; PKP OS in 2008 for pseudophakic bullous keratopathy on the eye with far advanced glaucoma. IOP was normal after previous filtering surgery. After PKP VA = 0,04; clear graft during 2 years; then gradually opacification and vascularization occurred. VA dropped to light perception. Echography OD — tumor h 8,29 mm, d 21,77 mm. No ingrowth of tumor into anterior segment of the eye; VA OD = 0 (no light perception. Concerning the need to enucleate the right eye with large choroidal melanoma, the advantage of using corneal autograft from OD to OS, location of tumor in the posterior pole with no ingrowth in anterior segment, the decision was made to perform the following operation — to enucleate the right eye and transplant simultaneously corneal graft from OD on OS. Patient was discharged from the Ophthalmology Hospital with VA OS = 0,01, during next week VA improved to 0,02. 8,0 mm graft isclear, fixed with 8 interruptured and continious suture 10 / 0‑nylon. Anterior chamber — normal depth, atrophic iris, stable position of PC IOL. Optic nerve head is pale with subtotal deep glaucomatous excavation. Conclusion: presented case report demonstrates the rarepossibility to use cornea after enucleation the eye with large malignant tumor (located in the posterior pole for grafting in the only eye with failed vascularised graft. It was the only possibility for this patient to restore some vision.

  19. Enucleaton of the right eye due to large choroidal melanoma with simultaneous penetrating cornea transplantation from OD to OS (Case report.

    Directory of Open Access Journals (Sweden)

    E. A. Korchuganova

    2014-07-01

    Full Text Available There is presentation case report of 75‑year old woman with choroidal melanoma (T3N0M0 on the right eye and failed graft on the left pseudophakic eye with far advanced glaucoma and ARMD. No treatment was given to the leading eye with VA 0,2. VA of OS = 1 / ∞ pr.l.certa; PKP OS in 2008 for pseudophakic bullous keratopathy on the eye with far advanced glaucoma. IOP was normal after previous filtering surgery. After PKP VA = 0,04; clear graft during 2 years; then gradually opacification and vascularization occurred. VA dropped to light perception. Echography OD — tumor h 8,29 mm, d 21,77 mm. No ingrowth of tumor into anterior segment of the eye; VA OD = 0 (no light perception. Concerning the need to enucleate the right eye with large choroidal melanoma, the advantage of using corneal autograft from OD to OS, location of tumor in the posterior pole with no ingrowth in anterior segment, the decision was made to perform the following operation — to enucleate the right eye and transplant simultaneously corneal graft from OD on OS. Patient was discharged from the Ophthalmology Hospital with VA OS = 0,01, during next week VA improved to 0,02. 8,0 mm graft isclear, fixed with 8 interruptured and continious suture 10 / 0‑nylon. Anterior chamber — normal depth, atrophic iris, stable position of PC IOL. Optic nerve head is pale with subtotal deep glaucomatous excavation. Conclusion: presented case report demonstrates the rarepossibility to use cornea after enucleation the eye with large malignant tumor (located in the posterior pole for grafting in the only eye with failed vascularised graft. It was the only possibility for this patient to restore some vision.

  20. Transplantation.

    Science.gov (United States)

    Faro, Albert; Weymann, Alexander

    2016-08-01

    Despite improvement in median life expectancy and overall health, some children with cystic fibrosis (CF) progress to end-stage lung or liver disease and become candidates for transplant. Transplants for children with CF hold the promise to extend and improve the quality of life, but barriers to successful long-term outcomes include shortage of suitable donor organs; potential complications from the surgical procedure and immunosuppressants; risk of rejection and infection; and the need for lifelong, strict adherence to a complex medical regimen. This article reviews the indications and complications of lung and liver transplantation in children with CF. PMID:27469184

  1. Establishment and characterization of a transplantable tumor line (RMM) and cell line (RMM-C) from a malignant amelanotic melanoma in the F344 rat, with particular reference to galectin-3 expression in vivo and in vitro.

    Science.gov (United States)

    Bondoc, Alexandra; Katou-Ichikawa, Chisa; Golbar, Hossain M; Tanaka, Miyuu; Izawa, Takeshi; Kuwamura, Mitsuru; Yamate, Jyoji

    2016-11-01

    To investigate characteristics of malignant melanomas with various pathobiological features, a homotransplantable tumor line (RMM) was established from a spontaneous amelanotic melanoma found in the pinna of an aged F344 rat. RMM tumors were transplanted in syngeneic rats by serial subcutaneous implantation with 100% intake. The original and RMM tumors consisted of spindle-shaped cells arranged mainly in interlacing bundles. Immunohistochemically, the neoplastic cells were positive to PNL-2 (melanocytes), nestin (neuroectodermal stem cells), S-100 (neurogenic cells) and vimentin (mesenchymal cells). Electron microscopically, tumor cells possessed single membrane-bound pre-melanosomes. Further, a cell line (RMM-C) was induced from an RMM tumor. RMM-C cells and the induced tumors in syngeneic rats showed immunohistochemical reactions similar to the original and RMM tumors. Interestingly, serum level of galectin-3 expression was increased with growing RMM tumors, and the expression was influenced by TNF-α (increase) or TGF-β1 (decrease), indicating a possible biomarker of amelanotic melanomas. The RMM tumors and RMM-C cell line could become useful tools for studies on the pathobiology, including tumor immunity, and development of therapeutic strategies against this malignancy. These tools are the first tumor lines of amelanotic melanomas in the rat. PMID:26949998

  2. Tumor vaccine strategies after allogeneic T-cell depleted bone marrow transplantation

    Directory of Open Access Journals (Sweden)

    Ferrara James L.M.

    2002-01-01

    Full Text Available Allogeneic bone marrow transplantation is currently restricted to hematological malignancies because of a lack of anti-tumor activity against solid cancers. We have tested a novel treatment strategy to stimulate specific anti-tumor activity against a solid tumor after transplantation by vaccination with irradiated tumor cells engineered to secrete granulocyte-macrophage colony-stimulating factor. Using the B16 melanoma model, we found that vaccination elicited potent anti-tumor activity in recipients of syngeneic bone marrow transplantation in a time dependent fashion, and that immune reconstitution was critical for the development of anti-tumor activity. Vaccination did not stimulate anti-tumor immunity after allogeneic bone marrow transplantation because of the post-transplantation immunodeficiency associated with graft-versus-host disease. Remarkably, vaccination was effective in stimulating potent and long-lasting anti-tumor activity in recipients of T cell-depleted allogeneic bone marrow. Thus T cells derived from donor stem cells were able to recognize tumor antigens even though they remained tolerant to host histocompatibility antigens. Donor leukocyte infusion from a donor immunized with the recipient-derived B16 vaccines enhanced clinical activity of tumor vaccines without exacerbating graft-versus-host disease and CD4+ T cells are essential for this enhancement. These results demonstrate that vaccination of both donors and recipients can stimulate potent anti-tumor effects without the induction of graft-versus-host disease, and this strategy has important implications for the treatment of patients with solid malignancies.

  3. TRANSPLANTATION

    Institute of Scientific and Technical Information of China (English)

    1996-01-01

    In order to reserch the influence of HLAmatch to the recovering of renal function afterrenal transplantation, the dare of HLA matchand uric RBP from 25 patients were collected.The results were shown that retinol-bindingprotein (RBP) was more sensitive than serumCr to reflect renal function. One monthpostoperation, the uric RBP value was less than

  4. TRANSPLANTATION

    Institute of Scientific and Technical Information of China (English)

    1996-01-01

    Objective: To explore the experience ofliver transpfantation in patients with terminalliver failure. Methods: From October 1991 toJuly 1995, 17 adults and 6 children underwentorthotopic liver transplantation. Preoperativediagnosis showed biliary atresia (n=5), Alagillesyndrome (n=1), primary biliary cirrhosis(n=2), cryptogenic cirrhosis (n=2), alcoholic

  5. Effects of Wnt-10b on proliferation and differentiation of murine melanoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Misu, Masayasu [Department of Pathogen, Infection and Immunity, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521 (Japan); Ouji, Yukiteru, E-mail: oujix@naramed-u.ac.jp [Department of Pathogen, Infection and Immunity, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521 (Japan); Kawai, Norikazu [Department of Pathogen, Infection and Immunity, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521 (Japan); Nishimura, Fumihiko [Department of Neurosurgery, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521 (Japan); Nakamura-Uchiyama, Fukumi [Department of Pathogen, Infection and Immunity, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521 (Japan); Yoshikawa, Masahide, E-mail: myoshika@naramed-u.ac.jp [Department of Pathogen, Infection and Immunity, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521 (Japan)

    2015-08-07

    In spite of the strong expression of Wnt-10b in melanomas, its role in melanoma cells has not been elucidated. In the present study, the biological effects of Wnt-10b on murine B16F10 (B16) melanoma cells were investigated using conditioned medium from Wnt-10b-producing COS cells (Wnt-CM). After 2 days of culture in the presence of Wnt-CM, proliferation of B16 melanoma cells was inhibited, whereas tyrosinase activity was increased. An in vitro wound healing assay demonstrated that migration of melanoma cells to the wound area was inhibited with the addition of Wnt-CM. Furthermore, evaluation of cellular senescence revealed prominent induction of SA-β-gal-positive senescent cells in cultures with Wnt-CM. Finally, the growth of B16 melanoma cell aggregates in collagen 3D-gel cultures was markedly suppressed in the presence of Wnt-CM. These results suggest that Wnt-10b represses tumor cell properties, such as proliferation and migration of B16 melanoma cells, driving them toward a more differentiated state along a melanocyte lineage. - Highlights: • Wnt-10b inhibited proliferation and migration of melanoma cells. • Wnt-10b induced tyrosinase activity and senescence of melanoma cells. • Wnt-10b suppressed growth of cell aggregates in collagen 3D-gel cultures. • Wnt-10b represses tumor cell properties, driving them toward a more differentiated state along a melanocyte lineage.

  6. Melanoma genetics

    DEFF Research Database (Denmark)

    Read, Jazlyn; Wadt, Karin A W; Hayward, Nicholas K

    2016-01-01

    Approximately 10% of melanoma cases report a relative affected with melanoma, and a positive family history is associated with an increased risk of developing melanoma. Although the majority of genetic alterations associated with melanoma development are somatic, the underlying presence of herita......Approximately 10% of melanoma cases report a relative affected with melanoma, and a positive family history is associated with an increased risk of developing melanoma. Although the majority of genetic alterations associated with melanoma development are somatic, the underlying presence...... in a combined total of approximately 50% of familial melanoma cases, the underlying genetic basis is unexplained for the remainder of high-density melanoma families. Aside from the possibility of extremely rare mutations in a few additional high penetrance genes yet to be discovered, this suggests a likely...... polygenic component to susceptibility, and a unique level of personal melanoma risk influenced by multiple low-risk alleles and genetic modifiers. In addition to conferring a risk of cutaneous melanoma, some 'melanoma' predisposition genes have been linked to other cancers, with cancer clustering observed...

  7. Lumican Inhibits SNAIL-Induced Melanoma Cell Migration Specifically by Blocking MMP-14 Activity

    Science.gov (United States)

    Stasiak, Marta; Boncela, Joanna; Perreau, Corinne; Karamanou, Konstantina; Chatron-Colliet, Aurore; Proult, Isabelle; Przygodzka, Patrycja; Chakravarti, Shukti; Maquart, François-Xavier; Kowalska, M. Anna; Wegrowski, Yanusz; Brézillon, Stéphane

    2016-01-01

    Lumican, a small leucine rich proteoglycan, inhibits MMP-14 activity and melanoma cell migration in vitro and in vivo. Snail triggers epithelial-mesenchymal transitions endowing epithelial cells with migratory and invasive properties during tumor progression. The aim of this work was to investigate lumican effects on MMP-14 activity and migration of Snail overexpressing B16F1 (Snail-B16F1) melanoma cells and HT-29 colon adenocarcinoma cells. Lumican inhibits the Snail induced MMP-14 activity in B16F1 but not in HT-29 cells. In Snail-B16F1 cells, lumican inhibits migration, growth, and melanoma primary tumor development. A lumican-based strategy targeting Snail-induced MMP-14 activity might be useful for melanoma treatment. PMID:26930497

  8. Electrochemotherapy by pulsed electromagnetic field treatment (PEMF in mouse melanoma B16F10 in vivo

    Directory of Open Access Journals (Sweden)

    Kranjc Simona

    2016-03-01

    Full Text Available Pulsed electromagnetic field (PEMF induces pulsed electric field, which presumably increases membrane permeabilization of the exposed cells, similar to the conventional electroporation. Thus, contactless PEMF could represent a promising approach for drug delivery.

  9. Asiaticoside, a component of Centella asiatica, inhibits melanogenesis in B16F10 mouse melanoma.

    Science.gov (United States)

    Kwon, Ku Jung; Bae, Seunghee; Kim, Karam; An, In Sook; Ahn, Kyu Joong; An, Sungkwan; Cha, Hwa Jun

    2014-07-01

    Melanogenesis is the process of generating pigmentation via melanin synthesis and delivery. Three key enzymes, tyrosinase, tyrosinase-related protein 1 (TRP1) and TRP2, metabolize melanin from L-tyrosine. Melanin synthesizing enzymes are regulated by microphthalmia-associated transcription factor (MITF). The titrated extract of Centella asiatica (TECA) contains the major components asiatic acid, asiaticoside and madecassic acid. The present study revealed that TECA reduces the melanin content in melanocytes. Moreover, the asiaticoside contained in TECA modulated melanogenesis by inhibiting tyrosinase mRNA expression. The decrease in tyrosinase mRNA levels was mediated through MITF. Uniquely, asiaticoside inhibited MITF by decreasing its DNA binding affinity. In conclusion, the results of the present study indicate that asiaticoside treatment may have beneficial effects in hyperpigmentation diseases or for skin whitening.

  10. The effect of antineoplastic drug NSC631570 on immunogenicity of B16 melanoma

    OpenAIRE

    Skivka, Larysa M.; Olexander G. Fedorchuk; Natalia O. Bezdeneznykh; Olexanrda O. Lykhova; Nadija I. Semesiuk; Yuri I. Kudryavets; Oksana M. Malanchuk

    2014-01-01

    Objectives: NSC631570 is a cytotoxic drug with the ability to be selectively accumulated in tumor tissue and activate apoptosis only in malignant cells and not in normal cells. Therapy with NSC631570 is accompanied by the stimulation of anticancer immune responses. It is known that cytotoxic anticancer drugs have additional effects on the immune system of tumor-bearing organism by increasing the immunogenic properties of tumor cells. This study aimed to investigate the immunogenicity of melan...

  11. Inhibitory activity of Bifidobacterium adolescent combined with cisplatin on melanoma in mice and its mechanism

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The aim of this study is to explore inhibitory activity of Bifidobacterium adolescent combined with cisplatin on the growth of melanoma(B16)in mice and the underlying mechanism.C57 mice were inoculated with B16 cancer cells to construct mouse model of melanoma and treated with bifidobacterium adolescent combined with cisplatin.Ratios of inhibitory activity on the growth of melanoma(B 16)were calculated.Pathology changes of the tumor were observed by HE staining.B 16 cell cycles were examined on a flow cytometer.Lymphocyte proliferation was measured with MTT assay and the T-cell subset was measured by double marked fluorescence.When bifidobacterium of 1010 cfu/L was injected,the ratio of inhibitory activity on the growth of melanoma(B16)reached 54%,which was similar to that of cisplatin group.The ratio of inhibitory activity reached 74.45% when the mice were treated by bifidobacterium combined with cisplatin,HE staining shows that bifidobacterium inhibited B16 cell proliferation and enhanced the cisplati(n)s killing activity on B16 cells.The results of flow cytometry demonstrated that B16 cell proliferation was arrested at G1 stage after treatment with bifidobacterium.The B16 cell proliferation was arrested at S stage after treatment with cisplatin.The CD4+ percentage increased and the difference was significant compared with the normal group after treatment with bifidobacterium,indicating that T-cell immune activity was enhanced.Treatment with bifidobacterium combined with cisplatin can enhance the inhibitory activity on the growth of melanoma(B16)of cisplatin.The mechanism of the inhibitory activity on B 16 cell proliferation is correlated with the enhanced immune activity in mice.

  12. Alphavirus replicon particles expressing TRP-2 provide potent therapeutic effect on melanoma through activation of humoral and cellular immunity.

    Directory of Open Access Journals (Sweden)

    Francesca Avogadri

    Full Text Available BACKGROUND: Malignant melanoma is the deadliest form of skin cancer and is refractory to conventional chemotherapy and radiotherapy. Therefore alternative approaches to treat this disease, such as immunotherapy, are needed. Melanoma vaccine design has mainly focused on targeting CD8+ T cells. Activation of effector CD8+ T cells has been achieved in patients, but provided limited clinical benefit, due to immune-escape mechanisms established by advanced tumors. We have previously shown that alphavirus-based virus-like replicon particles (VRP simultaneously activate strong cellular and humoral immunity against the weakly immunogenic melanoma differentiation antigen (MDA tyrosinase. Here we further investigate the antitumor effect and the immune mechanisms of VRP encoding different MDAs. METHODOLOGY/PRINCIPAL FINDINGS: VRP encoding different MDAs were screened for their ability to prevent the growth of the B16 mouse transplantable melanoma. The immunologic mechanisms of efficacy were investigated for the most effective vaccine identified, focusing on CD8+ T cells and humoral responses. To this end, ex vivo immune assays and transgenic mice lacking specific immune effector functions were used. The studies identified a potent therapeutic VRP vaccine, encoding tyrosinase related protein 2 (TRP-2, which provided a durable anti-tumor effect. The efficacy of VRP-TRP2 relies on a novel immune mechanism of action requiring the activation of both IgG and CD8+ T cell effector responses, and depends on signaling through activating Fcγ receptors. CONCLUSIONS/SIGNIFICANCE: This study identifies a VRP-based vaccine able to elicit humoral immunity against TRP-2, which plays a role in melanoma immunotherapy and synergizes with tumor-specific CD8+ T cell responses. These findings will aid in the rational design of future immunotherapy clinical trials.

  13. Role of SOCS-1 Gene on Melanoma Cell Growth and Tumor Development1

    OpenAIRE

    Scutti, Jorge A Borin; Matsuo, Alisson Leonardo; Pereira, Felipe Valença; Massaoka, Mariana Hiromi; Figueiredo, Carlos Rogério; Moreira, Dayson Friaça; Belizário, José Ernesto; Luiz R Travassos

    2011-01-01

    Melanoma is the most aggressive form of skin cancer, and its incidence has increased dramatically over the years. The murine B16F10 melanoma in syngeneic C57Bl/6 mice has been used as a highly aggressive model to investigate tumor development. Presently, we demonstrate in the B16F10-Nex2 subclone that silencing of SOCS-1, a negative regulator of Jak/Stat pathway, leads to reversal of the tumorigenic phenotype and inhibition of melanoma cell metastasis. SOCS-1 silencing with short hairpin RNA ...

  14. Cutaneous melanoma.

    Science.gov (United States)

    Eggermont, Alexander M M; Spatz, Alan; Robert, Caroline

    2014-03-01

    In the past decade, major advances have been made in the understanding of melanoma. New predisposition genes have been reported and key somatic events, such as BRAF mutation, directly translated into therapeutic management. Surgery for localised melanoma and regional lymph node metastases is the standard of care. Sentinel-node biopsy provides precise staging, but has not been reported to affect survival. The effect of lymph-node dissection on survival is a topic of investigation. Two distinct approaches have emerged to try to extend survival in patients with metastatic melanoma: immunomodulation with anti-CTLA4 monoclonal antibodies, and targeted therapy with BRAF inhibitors or MEK inhibitors for BRAF-mutated melanoma. The combination of BRAF inhibitors and MEK inhibitors might improve progression-free survival further and, possibly, increase overall survival. Response patterns differ substantially-anti-CTLA4 immunotherapy can induce long-term responses, but only in a few patients, whereas targeted drugs induce responses in most patients, but nearly all of them relapse because of pre-existing or acquired resistance. Thus, the long-term prognosis of metastatic melanoma remains poor. Anti-PD1 and anti-PDL1 antibodies have emerged as breakthrough drugs for melanoma that have high response rates and long durability. Biomarkers that have predictive value remain elusive in melanoma, although emerging data for adjuvant therapy indicate that interferon sensitivity is associated with ulceration of the primary melanoma. Intense investigation continues for clinical and biological markers that predict clinical benefit of immunotherapeutic drugs, such as interferon alfa or anti-CTLA4 antibodies, and the mechanisms that lead to resistance of targeted drugs.

  15. Up-regulation of hepatoma-derived growth factor facilitates tumor progression in malignant melanoma [corrected].

    Directory of Open Access Journals (Sweden)

    Han-En Tsai

    Full Text Available Cutaneous malignant melanoma is the fastest increasing malignancy in humans. Hepatoma-derived growth factor (HDGF is a novel growth factor identified from human hepatoma cell line. HDGF overexpression is correlated with poor prognosis in various types of cancer including melanoma. However, the underlying mechanism of HDGF overexpression in developing melanoma remains unclear. In this study, human melanoma cell lines (A375, A2058, MEL-RM and MM200 showed higher levels of HDGF gene expression, whereas human epidermal melanocytes (HEMn expressed less. Exogenous application of HDGF stimulated colony formation and invasion of human melanoma cells. Moreover, HDGF overexpression stimulated the degree of invasion and colony formation of B16-F10 melanoma cells whereas HDGF knockdown exerted opposite effects in vitro. To evaluate the effects of HDGF on tumour growth and metastasis in vivo, syngeneic mouse melanoma and metastatic melanoma models were performed by manipulating the gene expression of HDGF in melanoma cells. It was found that mice injected with HDGF-overexpressing melanoma cells had greater tumour growth and higher metastatic capability. In contrast, mice implanted with HDGF-depleted melanoma cells exhibited reduced tumor burden and lung metastasis. Histological analysis of excised tumors revealed higher degree of cell proliferation and neovascularization in HDGF-overexpressing melanoma. The present study provides evidence that HDGF promotes tumor progression of melanoma and targeting HDGF may constitute a novel strategy for the treatment of melanoma.

  16. Polysaccharide from Inonotus obliquus inhibits migration and invasion in B16-F10 cells by suppressing MMP-2 and MMP-9 via downregulation of NF-κB signaling pathway.

    Science.gov (United States)

    Lee, Ki Rim; Lee, Jong Seok; Kim, Young Rae; Song, In Gyu; Hong, Eock Kee

    2014-05-01

    Polysaccharides derived from Inonotus obliquus (PIO) are known to possess multiple pharmacological activities including antitumor activity. However, the possible molecular mechanisms of these activities are unknown. In the present study, we determined the anti-metastatic potential and signaling pathways of PIO in the highly metastatic B16-F10 mouse melanoma cell line in vitro. We found that PIO suppressed the migration and invasive ability of B16-F10 cells and decreased the expression levels and activities of matrix metalloproteinase (MMP)-2 and MMP-9. In addition, PIO decreased the phosphorylation levels of extracellular signal-regulated protein kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK); PIO also decreased the expression level of cyclooxygenase (COX)‑2 and inhibited the nuclear translocation of nuclear factor κB (NF-κB) in B16-F10 melanoma cells. These results suggest that PIO could suppress the invasion and migration of B16-F10 melanoma cells by reducing the expression levels and activities of MMP-2 and MMP-9 through suppressing MAPK, COX-2 and NF-κB signaling pathways. PMID:24677090

  17. Malignant Melanoma

    Directory of Open Access Journals (Sweden)

    Eshini Perera

    2013-12-01

    Full Text Available Melanomas are a major cause of premature death from cancer. The gradual decrease in rates of morbidity and mortality has occurred as a result of public health campaigns and improved rates of early diagnosis. Survival of melanoma has increased to over 90%. Management of melanoma involves a number of components: excision, tumor staging, re-excision with negative margins, adjuvant therapies (chemo, radiation or surgery, treatment of stage IV disease, follow-up examination for metastasis, lifestyle modification and counseling. Sentinel lymph node status is an important prognostic factor for survival in patients with a melanoma >1 mm. However, sentinel lymph node biopsies have received partial support due to the limited data regarding the survival advantage of complete lymph node dissection when a micrometastasis is detected in the lymph nodes. Functional mutations in the mitogen-activated pathways are commonly detected in melanomas and these influence the growth control. Therapies that target these pathways are rapidly emerging, and are being shown to increase survival rates in patients. Access to these newer agents can be gained by participation in clinical trials after referral to a multidisciplinary team for staging and re-excision of the scar.

  18. Bioelectric Applications for Treatment of Melanoma

    International Nuclear Information System (INIS)

    Two new cancer therapies apply bioelectric principles. These methods target tumor structures locally and function by applying millisecond electric fields to deliver plasmid DNA encoding cytokines using electrogene transfer (EGT) or by applying rapid rise-time nanosecond pulsed electric fields (nsPEFs). EGT has been used to locally deliver cytokines such as IL-12 to activate an immune response, resulting in bystander effects. NsPEFs locally induce apoptosis-like effects and affect vascular networks, both promoting tumor demise and restoration of normal vascular homeostasis. EGT with IL-12 is in melanoma clinical trials and nsPEFs are used in models with B16F10 melanoma in vitro and in mice. Applications of bioelectrics, using conventional electroporation and extensions of it, provide effective alternative therapies for melanoma

  19. Bioelectric Applications for Treatment of Melanoma

    Directory of Open Access Journals (Sweden)

    Richard Heller

    2010-09-01

    Full Text Available Two new cancer therapies apply bioelectric principles. These methods target tumor structures locally and function by applying millisecond electric fields to deliver plasmid DNA encoding cytokines using electrogene transfer (EGT or by applying rapid rise-time nanosecond pulsed electric fields (nsPEFs. EGT has been used to locally deliver cytokines such as IL-12 to activate an immune response, resulting in bystander effects. NsPEFs locally induce apoptosis-like effects and affect vascular networks, both promoting tumor demise and restoration of normal vascular homeostasis. EGT with IL-12 is in melanoma clinical trials and nsPEFs are used in models with B16F10 melanoma in vitro and in mice. Applications of bioelectrics, using conventional electroporation and extensions of it, provide effective alternative therapies for melanoma.

  20. Bioelectric Applications for Treatment of Melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Beebe, Stephen J., E-mail: sbeebe@odu.edu; Schoenbach, Karl H.; Heller, Richard [Frank Reidy Research Center for Bioelectrics/Old Dominion University 4211 Monarch Way, Suite 300, Norfolk, Virginia 23508 (United States)

    2010-09-27

    Two new cancer therapies apply bioelectric principles. These methods target tumor structures locally and function by applying millisecond electric fields to deliver plasmid DNA encoding cytokines using electrogene transfer (EGT) or by applying rapid rise-time nanosecond pulsed electric fields (nsPEFs). EGT has been used to locally deliver cytokines such as IL-12 to activate an immune response, resulting in bystander effects. NsPEFs locally induce apoptosis-like effects and affect vascular networks, both promoting tumor demise and restoration of normal vascular homeostasis. EGT with IL-12 is in melanoma clinical trials and nsPEFs are used in models with B16F10 melanoma in vitro and in mice. Applications of bioelectrics, using conventional electroporation and extensions of it, provide effective alternative therapies for melanoma.

  1. The kin17 Protein in Murine Melanoma Cells

    Directory of Open Access Journals (Sweden)

    Anelise C. Ramos

    2015-11-01

    Full Text Available kin17 has been described as a protein involved in the processes of DNA replication initiation, DNA recombination, and DNA repair. kin17 has been studied as a potential molecular marker of breast cancer. This work reports the detection and localization of this protein in the murine melanoma cell line B16F10-Nex2 and in two derived subclones with different metastatic potential, B16-8HR and B16-10CR. Nuclear and chromatin-associated protein fractions were analyzed, and kin17 was detected in all fractions, with an elevated concentration observed in the chromatin-associated fraction of the clone with low metastatic potential, suggesting that the kin17 expression level could be a marker of melanoma.

  2. Melanoma immunotherapy.

    Science.gov (United States)

    Sivendran, Shanthi; Glodny, Bradley; Pan, Michael; Merad, Miriam; Saenger, Yvonne

    2010-01-01

    Melanoma immunotherapy has been an area of intense research for decades, and this work is now yielding more tangible results for patients. Work has focused on 4 main areas: cytokine therapy, administration of immune-modulating antibodies, adoptive T-cell therapy, and vaccines. Cytokine therapy is an established treatment for advanced melanoma, and immune-modulating antibodies have recently emerged as an exciting new area of drug development with efficacy now established in a phase III trial. Adoptive T-cell therapy provides the proof of principle that T cells can attack and eliminate tumors. It has been challenging, however, to adapt this treatment for widespread use. Vaccines have generally yielded poor results, but intratumor pathogen-based strategies have shown encouraging results in recent trials, perhaps due to stronger immune stimulation. A review of the field of melanoma immunotherapy is provided here, with emphasis on those agents that have reached clinical testing. Novel strategies to induce the immune system to attack melanomas are reviewed. In the future, it is envisioned that immunotherapy will have further application in combination with cytotoxic and targeted therapies.

  3. The chemotherapeutic effect of essential oil of Plectranthus amboinicus (Lour) on lung metastasis developed by B16F-10 cell line in C57BL/6 mice.

    Science.gov (United States)

    Manjamalai, A; Grace, V M Berlin

    2013-01-01

    Current investigation is to evaluate the anticancer activity of the essential oil of Plectranthus amboinicus (Lour) on B16F-10 melanoma cell line injected C57BL/6 mice, and it was simultaneously treated with the essential oil of P. amboinicus (Lour) (50 μg/dose) via i.p. for 21 days. The present investigation exhibited the potent chemotherapeutic/chemopreventive effect of the essential oil of P. amboinicus (Lour) over lung metastasis that developed. To our knowledge, this is the first report in evaluating the effect of essential oil of P. amboinicus (Lour) using lung cancer model. PMID:23249189

  4. Gallium-67-Labeled Lactam Bridge-Cyclized Alpha-Melanocyte Stimulating Hormone Peptide for Primary and Metastatic Melanoma Imaging

    OpenAIRE

    Guo, Haixun; Yang,Jianquan; Shenoy, Nalini; Miao, Yubin

    2009-01-01

    The purpose of this study was to examine the melanoma imaging properties of a novel 67Ga-labeled lactam bridge-cyclized alpha-melanocyte stimulating hormone (α-MSH) peptide. A lactam bridge-cyclized α-MSH peptide, DOTA-GlyGlu-CycMSH {DOTA-Gly-Glu-c[Lys-Nle-Glu-His-DPhe-Arg-Trp-Gly-Arg-Pro-Val-Asp]}, was synthesized and radiolabeled with 67Ga. The melanoma targeting and pharmacokinetic properties of 67Ga-DOTA-GlyGlu-CycMSH were determined in B16/F1 flank primary melanoma-bearing and B16/F10 pu...

  5. The application of quantum dots for the melanoma tumor in vivo imaging

    Science.gov (United States)

    Feng, Yayi; Zhai, Peng; Wang, Xiaomei; Ying, Ming; Wu, Jinbo; Zhu, Xiaomei; Lin, Guimiao; Chen, Qiang; Xu, Gaixia

    2014-09-01

    Objective: Over the past decade, fluorescent semiconductor nanocrystals, also known as quantum dots (QDs), have been applied in biomedical imaging in vitro and in vivo because of their fascinating optical properties. In this work, we investigated the application of CdTe QDs for tumor fluorescence in vivo imaging. Methods: The transparent dorsal skin fold window chamber (DSFC) was constructed on the 4~6 week-old BALB/c mice. The melanoma cells stably expressing green fluorescent protein ---ZsGreen were transplanted into the chamber and the melanoma DSFC model was established successfully. The water soluble CdTe QDs were synthesized and then administrated in the model through the tail intravenous injection. The fluorescent expression of B16 cells were assayed by fluorescent microscopy, the tumor growth, the blood capillaries distributions and its dynamic changes were observed by stereomicroscopy and laser scanning confocal microscopy. Results: The results demonstrated that the expression efficiency of ZsGreen was 41%, which met the experimental requirement. The tumors was visible inside the chamber after implantation of melanoma cells for 5~6 days, while no obvious changes in mice behaviors were found. After injection of the QDs, CdTe QDs accumulated at the invading edge of a range of solid tumor. We could also observe the tumor cells growth near the blood vessels, the angiogenesis occurred inside the tumor and the local blood capillaries increased. Conclusions: This work provided a new strategy for the tumor in vivo imaging and the development of targeted antineoplastic drugs.

  6. Melanoma cells treated with GGTI and IFN-gamma allow murine vaccination and enhance cytotoxic response against human melanoma cells.

    Directory of Open Access Journals (Sweden)

    Guillaume Sarrabayrouse

    Full Text Available BACKGROUND: Suboptimal activation of T lymphocytes by melanoma cells is often due to the defective expression of class I major histocompatibility antigens (MHC-I and costimulatory molecules. We have previously shown that geranylgeranyl transferase inhibition (done with GGTI-298 stimulates anti-melanoma immune response through MHC-I and costimulatory molecule expression in the B16F10 murine model [1]. METHODOLOGY/PRINCIPAL FINDINGS: In this study, it is shown that vaccination with mIFN-gand GGTI-298 pretreated B16F10 cells induces a protection against untreated tumor growth and pulmonary metastases implantation. Furthermore, using a human melanoma model (LB1319-MEL, we demonstrated that in vitro treatment with hIFN-gamma and GGTI-298 led to the up regulation of MHC-I and a costimulatory molecule CD86 and down regulation of an inhibitory molecule PD-1L. Co-culture experiments with peripheral blood mononuclear cells (PBMC revealed that modifications induced by hIFN-gamma and GGTI-298 on the selected melanoma cells, enables the stimulation of lymphocytes from HLA compatible healthy donors. Indeed, as compared with untreated melanoma cells, pretreatment with hIFN-gamma and GGTI-298 together rendered the melanoma cells more efficient at inducing the: i activation of CD8 T lymphocytes (CD8+/CD69+; ii proliferation of tumor-specific CD8 T cells (MelanA-MART1/TCR+; iii secretion of hIFN-gamma; and iv anti-melanoma specific cytotoxic cells. CONCLUSIONS/SIGNIFICANCE: These data indicate that pharmacological treatment of melanoma cell lines with IFN-gamma and GGTI-298 stimulates their immunogenicity and could be a novel approach to produce tumor cells suitable for vaccination and for stimulation of anti-melanoma effector cells.

  7. Adjuvant therapy of melanoma.

    Science.gov (United States)

    Agarwala, S S; Kirkwood, J M

    1998-06-01

    Patients with AJCC Stage IIB and III melanoma have a poor 5-year survival rate which has been the driving force behind attempts to find an effective adjuvant therapy for this stage of disease that would effectively reduce relapse and improve survival. Immunotherapy with bacillus Calmette-Guerin (BCG), Corynebacterium parvum, and levamisole have not been successful in achieving this goal, nor have trials with chemotherapy in the adjuvant setting, including high-dose chemotherapy with autologous bone marrow transplantation. The recent Eastern Cooperative Oncology Group (ECOG) 1684 study showed significant improvement in relapse-free and overall survival with high doses of alpha interferon (IFNalpha) given for 1 year. Lower dosages of IFNalpha have to date been unsuccessful in impacting upon long-term survival. Recent data with vaccines have been encouraging, and the GM2-KLH vaccine is the focus of ongoing intergroup study comparing this treatment with IFNalpha in resected Stage IIB and III melanoma. The various regimens are reviewed in this article. PMID:9588723

  8. Choroidal melanoma

    International Nuclear Information System (INIS)

    A useful and practical guide is developed to better track to the uveal melanoma, due to its highly malignant character. Melanoma of the uveal tract (choroid, iris, ciliary body) has been the intraocular tumor most frequent in adults. The biopsy has been inaccessible, due to its location; therefore, the diagnostic should be based on clinical examination and the correct utilization of the diagnostic procedures (ultrasound, fluorescent angiography, computed axial tomography and magnetic resonance). The cases are diagnosed in the histological examination of the operatory piece post-enucleation for other causes. Epidemiological research has been key to determine the associated factors and better to understand the mechanisms of onset of the disease. Anatomopathological studies of choroidal melanoma have permitted to know the natural history of the disease. The decrease of the visual acuity, pain or inflammation are presented as a defect in the visual field. Different techniques to diagnose the disease are explained. Ultrasound in mode A and B, computed axial tomography and magnetic resonance are the diagnostic method of election. Ultrasound has been the primary method of diagnostic, giving the size and vascularisation, useful in tracking, when they are treated in shape conservatively, showing changes in echogenicity and less vascularisation as good response to treatment. The treatments of choroidal melanoma are specified. The correct interpretation of the clinical symptoms and early utilization of diagnostic imaging methods, have permitted to establish the adequate therapeutic and to avoid local and distant metastasis. The uveal melanoma, depending on their size and location, traditionally has been treated by enucleation. Data from the literature and authors, have promoted the conservation of the ocular globe, depending on the size of the tumor. Transpupillary thermotherapy has been an available alternative for small tumors in Costa Rica and level of social security

  9. Nutrition and melanoma prevention.

    Science.gov (United States)

    Jensen, J Daniel; Wing, Gregory J; Dellavalle, Robert P

    2010-01-01

    Melanoma has continued to rise in incidence despite public efforts to promote sun protection behaviors. Because sunscreen use does not completely prevent skin cancer induced by ultraviolet radiation, additional chemopreventive methods for protecting against and reversing the effects of ultraviolet photodamage need evaluation. Recent years have brought increased interest in dietary factors, such as natural botanicals and vitamins, for the prevention of melanoma. This contribution provides a narrative review of the relevant, nutrition-related literature found by searching the keywords "melanoma chemoprevention," "nutrition and melanoma," "dietary botanicals and melanoma prevention," "green tea and melanoma," "vitamin D and melanoma," and "vitamin E and melanoma" in the PubMed database. Although randomized controlled trials of humans are lacking, basic science and epidemiologic studies show promising benefits of many natural products in chemoprevention for melanoma. Future studies, hopefully, will yield concrete answers and clarify the role of commonly available dietary nutrients in melanoma chemoprevention.

  10. Cutaneous Melanoma in Asians.

    Science.gov (United States)

    Kim, Sang Yub; Yun, Sook Jung

    2016-09-01

    Malignant melanoma is a rare disease in Asians but potentially the most aggressive form of skin cancer worldwide. It can occur in any melanocyte-containing anatomic site. Four main cutaneous melanoma subtypes are recognized: lentigo maligna melanoma, superficial spreading melanoma, acral lentiginous melanoma (ALM), and nodular melanoma. Generally, excessive exposure to ultraviolet (UV) radiation increases the risk of melanoma. The exception is ALM, which is the most common melanoma subtype in Asians and is not associated with UV radiation. ALM presents as dark brownish to black, irregular maculopatches, nodules, or ulcers on the palms, soles, and nails. The lesions may be misdiagnosed as more benign lesions, such as warts, ulcers, hematomas, foreign bodies, or fungal infections, especially in amelanotic acral melanomas where black pigments are absent. The aim of this brief review is to improve understanding and the rate of early detection thereby reducing mortality, especially regarding cutaneous melanoma in Asians. PMID:27689028

  11. Ethanolic Extract of Algerian Propolis and Galangin Decreased Murine Melanoma T.

    Science.gov (United States)

    Benguedouar, Lamia; Lahouel, Mesbah; Gangloff, Sophie C; Durlach, Anne; Grange, Florent; Bernard, Philippe; Antonicelli, Frank

    2016-01-01

    Melanoma is the more dangerous skin cancer, and metastatic melanoma still carries poor prognosis. Despite recent therapeutic advances, prolonged survival remains rare and research is still required. Propolis extracts from many countries have attracted a great deal of attention for their biological properties. We here investigated the ability of an ethanolic extract of Algerian propolis (EEP) to control melanoma tumour growth when given to mice bearing B16F1melanoma tumour either as preventive or as therapeutic treatment. EEP given after tumour occurrence increased mice survival (+30%) and reduced tumour growth (-75%). This was associated with a decrease of the Mitotic Index (-75%) and of Ki-67 (-50%) expression. When given either before or both before and after tumour occurrence, EEP reduced tumour growth but without prolonging mice life. Isolation of B16F1 melanoma cells from resected tumour showed that preventive and curative EEP treatments reduced invasiveness by 55% and 40% respectively compared to control. Galangin, one of the most abundant flavonoids in propolis, significantly reduced the number of melanoma cells in vitro and induced autophagy/apoptosis dose dependently. In conclusion, we showed that EEP reduced melanoma tumour progression/dissemination and could extend mice lifespan when used as therapeutic treatment. Then, EEP may help patients with melanoma when used as a complementary therapy to classical treatment for which autophagy is not contraindicated. PMID:26863880

  12. Recombinant Interferon Alfa-2b in Treating Patients With Melanoma

    Science.gov (United States)

    2016-05-17

    Stage IA Skin Melanoma; Stage IB Skin Melanoma; Stage IIA Skin Melanoma; Stage IIB Skin Melanoma; Stage IIC Skin Melanoma; Stage IIIA Skin Melanoma; Stage IIIB Skin Melanoma; Stage IIIC Skin Melanoma; Stage IV Skin Melanoma

  13. What Is Melanoma Skin Cancer?

    Science.gov (United States)

    ... statistics for melanoma skin cancer What is melanoma skin cancer? Cancer starts when cells in the body begin ... causing the skin to tan or darken. Melanoma skin cancers Melanoma is a cancer that begins in the ...

  14. Recombinant adenovirus snake venom cystatin inhibits the growth, invasion, and metastasis of B16F10 cells in vitro and in vivo.

    Science.gov (United States)

    Xie, Qun; Tang, Nanhong; Lin, Yangyuan; Wang, Xiaoqian; Lin, Xu; Lin, Jianyin

    2013-12-01

    Previous studies have shown that transfection of the snake venom cystatin (sv-cystatin) gene can inhibit the invasion and metastasis of tumor cells. The aim of this study was to investigate the pharmaceutical applications of sv-cystatin in melanoma gene therapy. We constructed a recombinant adenovirus carrying sv-cystatin (Ad/sv-cystatin) and a control virus (Ad/null). Matrigel assays were used to assess melanoma cell migration and invasiveness in vitro. The antimelanoma effects of Ad/sv-cystatin were assessed in a syngeneic mouse model with an experimental lung colonization assay. Ad/sv-cystatin significantly inhibited the invasion and growth of B16F10 cells in vitro compared with control and Ad/null. Ad/sv-cystatin significantly inhibited experimental lung colonization in C57BL/6 mice as compared with that in control (Pcystatin slowed the increase in lung weight in C57BL/6 mice as compared with that in control mice (Pcystatin suppresses mouse melanoma invasion, metastasis, and growth in vitro and in vivo. Our findings provide support for the further examination of the pharmaceutical applications of Ad/sv-cystatin.

  15. Transfer of the human sodium/iodide symporter gene enhances iodide uptake in melanoma cells

    International Nuclear Information System (INIS)

    Objective: To obtain human sodium/iodide symporter (hNIS) cDNA and to study its biological property and potential use as a therapeutic radioiodide for melanoma. Methods: hNIS gene cDNA was amplified with total RNA from human thyroid tissue by RT-PCR. The hNIS cDNA was inserted into cloning vector pUCm-T and subcloned into eukaryotic expression vector pcDNA3. The recombinant plasmid pcDNA3-hNIS was introduced into B16 cells using the electroporation technique. The uptake and efflux of iodide was examined in vitro. Results: The cloned hNIS cDNA sequence was identical to the published sequence. Two novel cell lines named B16-A containing hNIS and B16-B containing pcDNA3 only were established. The resultant cell line B16-A accumulated 17 and 19 times more radioiodide in vitro than B16 and B16-B did, respectively. However the efflux of iodide from B16-A was also rapid ( T1/2=10 min). Conclusions: Our preliminary data indicate that the transduction of the hNIS gene per se is sufficient to induce iodide transport in melanoma cells in vitro, but its T1/2 is short. With regard to therapeutic application, however, further investigation is necessary so as to develop a method of maintaining more radioiodide in the cells for long enough to produce greater therapeutic effects

  16. Cripto-1 vaccination elicits protective immunity against metastatic melanoma.

    Science.gov (United States)

    Ligtenberg, M A; Witt, K; Galvez-Cancino, F; Sette, A; Lundqvist, A; Lladser, A; Kiessling, R

    2016-05-01

    Metastatic melanoma is a fatal disease that responds poorly to classical treatments but can be targeted by T cell-based immunotherapy. Cancer vaccines have the potential to generate long-lasting cytotoxic CD8(+) T cell responses able to eradicate established and disseminated tumors. Vaccination against antigens expressed by tumor cells with enhanced metastatic potential represents a highly attractive strategy to efficiently target deadly metastatic disease. Cripto-1 is frequently over-expressed in human carcinomas and melanomas, but is expressed only at low levels on normal differentiated tissues. Cripto-1 is particularly upregulated in cancer-initiating cells and is involved in cellular processes such as cell migration, invasion and epithelial-mesenchymal transition, which are hallmarks of aggressive cancer cells able to initiate metastatic disease. Here, we explored the potential of Cripto-1 vaccination to target metastatic melanoma in a preclinical model. Cripto-1 was overexpressed in highly metastatic B16F10 cells as compared to poorly metastatic B16F1 cells. Moreover, B16F10 cells grown in sphere conditions to enrich for cancer stem cells (CSC) progressively upregulated cripto1 expression. Vaccination of C57Bl/6 mice with a DNA vaccine encoding mouse Cripto-1 elicited a readily detectable/strong cytotoxic CD8(+) T cell response specific for a H-2 Kb-restricted epitope identified based on its ability to bind H-2(b) molecules. Remarkably, Cripto-1 vaccination elicited a protective response against lung metastasis and subcutaneous challenges with highly metastatic B16F10 melanoma cells. Our data indicate that vaccination against Cripto-1 represents a novel strategy to be tested in the clinic. PMID:27467944

  17. Liposomal encapsulation enhances the antitumour efficacy of the vascular disrupting agent ZD6126 in murine B16.F10 melanoma

    OpenAIRE

    Fens, M H A M; Hill, K. J.; Issa, J; Ashton, S E; Westwood, F. R.; Blakey, D C; Storm, G; Ryan, A J; Schiffelers, R.M.

    2008-01-01

    Vascular disrupting agents (VDAs) are able to affect selectively tumour endothelial cell morphology resulting in vessel occlusion and widespread tumour cell necrosis. However, single-agent antitumour activity of VDAs is typically limited, as tumour regrowth occurs rapidly following drug treatment. To improve the therapeutic effectiveness of VDAs, we investigated liposomal targeting using ZD6126 as a model VDA. ZD6126 is a phosphate-prodrug of the tubulin-binding vascular disrupting agent ZD61...

  18. The use of anchored agonists of phagocytic receptors for cancer immunotherapy: B16-F10 murine melanoma model.

    Directory of Open Access Journals (Sweden)

    Tereza Janotová

    Full Text Available The application of the phagocytic receptor agonists in cancer immunotherapy was studied. Agonists (laminarin, molecules with terminal mannose, N-Formyl-methioninyl-leucyl-phenylalanine were firmly anchored to the tumor cell surface. When particular agonists of phagocytic receptors were used together with LPS (Toll-like receptor agonist, high synergy causing tumour shrinkage and a temporary or permanent disappearance was observed. Methods of anchoring phagocytic receptor agonists (charge interactions, anchoring based on hydrophobic chains, covalent bonds and various regimes of phagocytic agonist/LPS mixture applications were tested to achieve maximum therapeutic effect. Combinations of mannan/LPS and f-MLF/LPS (hydrophobic anchors in appropriate (pulse regimes resulted in an 80% and 60% recovery for mice, respectively. We propose that substantial synergy between agonists of phagocytic and Toll-like receptors (TLR is based on two events. The TLR ligand induces early and massive inflammatory infiltration of tumors. The effect of this cell infiltrate is directed towards tumor cells, bearing agonists of phagocytic receptors on their surface. The result of these processes was effective killing of tumor cells. This novel approach represents exploitation of innate immunity mechanisms for treating cancer.

  19. Inhibitory and Acceleratory Effects of Inonotus obliquus on Tyrosinase Activity and Melanin Formation in B16 Melanoma Cells

    OpenAIRE

    Zheng-Fei Yan; Yang Yang; Feng-Hua Tian; Xin-Xin Mao; Yu Li; Chang-Tian Li

    2014-01-01

    The aim of the present study is to preliminarily investigate the antimelanogenesis effect of Inonotus obliquus extracts by cell-free mushroom tyrosinase assay. It was found that petroleum ether and n-butanol extracts might contain unknown potential tyrosinase inhibitors, while its ethyl acetate extract might contain some unknown accelerators. Six compounds were isolated and their structures were identified by interpretation of NMR data and nicotinic acid was first discovered in Inonotus obliq...

  20. Inhibitory and Acceleratory Effects of Inonotus obliquus on Tyrosinase Activity and Melanin Formation in B16 Melanoma Cells.

    Science.gov (United States)

    Yan, Zheng-Fei; Yang, Yang; Tian, Feng-Hua; Mao, Xin-Xin; Li, Yu; Li, Chang-Tian

    2014-01-01

    The aim of the present study is to preliminarily investigate the antimelanogenesis effect of Inonotus obliquus extracts by cell-free mushroom tyrosinase assay. It was found that petroleum ether and n-butanol extracts might contain unknown potential tyrosinase inhibitors, while its ethyl acetate extract might contain some unknown accelerators. Six compounds were isolated and their structures were identified by interpretation of NMR data and nicotinic acid was first discovered in Inonotus obliquus. In cells testing, betulin and trametenolic acid decreased tyrosinase activity and melanin content, while inotodiol and lanosterol significantly increased tyrosinase activity and melanin content, showing an AC⁡50 of 9.74 and 8.43 μM, respectively. Nicotinie acid, 3β,22,25-trihydroxy-lanosta-8-ene, had a little or no effect on tyrosinase. Betulin exhibited a mode of noncompetitive inhibition with a K I = K IS of 0.4 μM on tyrosinase activity showing an IC50 of 5.13 μM and being more effective than kojic acid (6.43 μM), and trametenolic acid exhibited a mode of mixed inhibition with a K I of 0.9 μM, K IS of 0.5 μM, and an IC50 of 7.25 μM. We proposed betulin and trametenolic acid as a new candidate of potent tyrosinase inhibitors and inotodiol and lanosterol as accelerators that could be used as therapeutic agent. PMID:25197307

  1. Suppressive Effect of Juzen-Taiho-To on Lung Metastasis of B16 Melanoma Cells In Vivo

    OpenAIRE

    Takako Matsuda; Katsuhiko Maekawa; Kazuhito Asano; Tadashi Hisamitsu

    2011-01-01

    Juzen-Taiho-To (JTT) is well known to be one of Kampo (Japanese herbal) medicine consisted of 10 component herbs and used for the supplemental therapy of cancer patients with remarkably success. However, the precise mechanisms by which JTT could favorably modify the clinical conditions of cancer patients are not well defined. The present study, therefore, was undertaken to examine the possible mechanisms of JTT on prevention of cancer metastasis using experimental mouse model. JTT was well mi...

  2. Inhibitory and Acceleratory Effects of Inonotus obliquus on Tyrosinase Activity and Melanin Formation in B16 Melanoma Cells

    Directory of Open Access Journals (Sweden)

    Zheng-Fei Yan

    2014-01-01

    Full Text Available The aim of the present study is to preliminarily investigate the antimelanogenesis effect of Inonotus obliquus extracts by cell-free mushroom tyrosinase assay. It was found that petroleum ether and n-butanol extracts might contain unknown potential tyrosinase inhibitors, while its ethyl acetate extract might contain some unknown accelerators. Six compounds were isolated and their structures were identified by interpretation of NMR data and nicotinic acid was first discovered in Inonotus obliquus. In cells testing, betulin and trametenolic acid decreased tyrosinase activity and melanin content, while inotodiol and lanosterol significantly increased tyrosinase activity and melanin content, showing an AC⁡50 of 9.74 and 8.43 μM, respectively. Nicotinie acid, 3β,22,25-trihydroxy-lanosta-8-ene, had a little or no effect on tyrosinase. Betulin exhibited a mode of noncompetitive inhibition with a KI=KIS of 0.4 μM on tyrosinase activity showing an IC50 of 5.13 μM and being more effective than kojic acid (6.43 μM, and trametenolic acid exhibited a mode of mixed inhibition with a KI of 0.9 μM, KIS of 0.5 μM, and an IC50 of 7.25 μM. We proposed betulin and trametenolic acid as a new candidate of potent tyrosinase inhibitors and inotodiol and lanosterol as accelerators that could be used as therapeutic agent.

  3. Cinnamomum cassia Essential Oil Inhibits α-MSH-Induced Melanin Production and Oxidative Stress in Murine B16 Melanoma Cells

    OpenAIRE

    Ying Shih; Shih-Lan Hsu; Yu-Che Lin; Chen-Tien Chang; Su-Tze Chou; Wen-Lun Chang

    2013-01-01

    Essential oils extracted from aromatic plants exhibit important biological activities and have become increasingly important for the development of aromatherapy for complementary and alternative medicine. The essential oil extracted from Cinnamomum cassia Presl (CC-EO) has various functional properties; however, little information is available regarding its anti-tyrosinase and anti-melanogenic activities. In this study, 16 compounds in the CC-EO have been identified; the major components of t...

  4. Thymoquinone suppresses metastasis of melanoma cells by inhibition of NLRP3 inflammasome

    Energy Technology Data Exchange (ETDEWEB)

    Ahmad, Israr; Muneer, Kashiff M.; Tamimi, Iman A.; Chang, Michelle E.; Ata, Muhammad O. [Department of Dermatology and Skin Diseases Research Center, University of Alabama at Birmingham, AL (United States); Yusuf, Nabiha, E-mail: nabiha@uab.edu [Department of Dermatology and Skin Diseases Research Center, University of Alabama at Birmingham, AL (United States); Veteran Affairs Medical Center, Birmingham, University of Alabama at Birmingham, AL (United States); Comprehensive Cancer Center, University of Alabama at Birmingham, AL (United States)

    2013-07-01

    The inflammasome is a multi-protein complex which when activated regulates caspase-1 activation and IL-1β and IL-18 secretion. The NLRP3 (NACHT, LRR, and pyrin domain-containing protein 3) inflammasome is constitutively assembled and activated in human melanoma cells. We have examined the inhibitory effect of thymoquinone (2-isopropyl-5-methylbenzo-1,4-quinone), a major ingredient of black seed obtained from the plant Nigella sativa on metastatic human (A375) and mouse (B16F10) melanoma cell lines. We have assessed whether thymoquinone inhibits metastasis of melanoma cells by targeting NLRP3 subunit of inflammasomes. Using an in vitro cell migration assay, we found that thymoquinone inhibited the migration of both human and mouse melanoma cells. The inhibitory effect of thymoquinone on metastasis was also observed in vivo in B16F10 mouse melanoma model. The inhibition of migration of melanoma cells by thymoquinone was accompanied by a decrease in expression of NLRP3 inflammasome resulting in decrease in proteolytic cleavage of caspase-1. Inactivation of caspase-1 by thymoquinone resulted in inhibition of IL-1β and IL-18. Treatment of mouse melanoma cells with thymoquinone also inhibited NF-κB activity. Furthermore, inhibition of reactive oxygen species (ROS) by thymoquinone resulted in partial inactivation of NLRP3 inflammasome. Thus, thymoquinone exerts its inhibitory effect on migration of human and mouse melanoma cells by inhibition of NLRP3 inflammasome. Thus, our results indicate that thymoquinone can be a potential immunotherapeutic agent not only as an adjuvant therapy for melanoma, but also, in the control and prevention of metastatic melanoma. - Highlights: • Thymoquinone causes inhibition of migration of melanoma cells. • Thymoquinone causes inhibition of metastasis in vivo. • Thymoquinone causes inhibition of migration by activation of NLRP3 inflammasome.

  5. Improved anti-melanoma effect of a transdermal mitoxantrone ethosome gel.

    Science.gov (United States)

    Yu, Xiang; Du, Lina; Li, Yu; Fu, Guiying; Jin, Yiguang

    2015-07-01

    Melanomas are malignant tumors characterized by early metastasis, rapid development, poor prognosis and high mortality. A highly effective and convenient method is necessary for long-term treatment of melanomas. Mitoxantrone (MTO) was topically applied for melanoma therapy using an MTO ethosome gel. Firstly, an ethosome was prepared from MTO, phospholipids, ethanol and water followed by addition of hydroxypropyl methylcellulose to obtain an ethosome gel. The ethosome was characterized. The cytotoxicity on B16 melanoma cells was evaluated on an electrical cell-substrate impedance sensing system with a novel modified chip. In vivo anti-melanoma effect of the ethosome gel was explored. Immunohistochemical and flow cytometric investigations were done. The MTO ethosomes had the size of 78nm and the zeta potential of -55mV. The ethosomes were flexible vesicles and showed much higher in vitro permeability across the rat skin than MTO aqueous solutions. The ethosomes had significant cytotoxicity and higher in vivo anti-melanoma effect than MTO solutions. The calreticulin membrane translocation of B16 cells was improved by the MTO ethosomes and the cell uptake of MTO was confirmed. The MTO ethosome gel is a promising transdermal delivery system for melanoma therapy with the advantages of non-invasion and no significant side effects. PMID:26211575

  6. Improved anti-melanoma effect of a transdermal mitoxantrone ethosome gel.

    Science.gov (United States)

    Yu, Xiang; Du, Lina; Li, Yu; Fu, Guiying; Jin, Yiguang

    2015-07-01

    Melanomas are malignant tumors characterized by early metastasis, rapid development, poor prognosis and high mortality. A highly effective and convenient method is necessary for long-term treatment of melanomas. Mitoxantrone (MTO) was topically applied for melanoma therapy using an MTO ethosome gel. Firstly, an ethosome was prepared from MTO, phospholipids, ethanol and water followed by addition of hydroxypropyl methylcellulose to obtain an ethosome gel. The ethosome was characterized. The cytotoxicity on B16 melanoma cells was evaluated on an electrical cell-substrate impedance sensing system with a novel modified chip. In vivo anti-melanoma effect of the ethosome gel was explored. Immunohistochemical and flow cytometric investigations were done. The MTO ethosomes had the size of 78nm and the zeta potential of -55mV. The ethosomes were flexible vesicles and showed much higher in vitro permeability across the rat skin than MTO aqueous solutions. The ethosomes had significant cytotoxicity and higher in vivo anti-melanoma effect than MTO solutions. The calreticulin membrane translocation of B16 cells was improved by the MTO ethosomes and the cell uptake of MTO was confirmed. The MTO ethosome gel is a promising transdermal delivery system for melanoma therapy with the advantages of non-invasion and no significant side effects.

  7. General Information about Melanoma

    Science.gov (United States)

    ... Screening Research Melanoma Treatment (PDQ®)–Patient Version General Information About Melanoma Go to Health Professional Version Key ... the PDQ Adult Treatment Editorial Board . Clinical Trial Information A clinical trial is a study to answer ...

  8. Treatment Option Overview (Melanoma)

    Science.gov (United States)

    ... of Skin Cancer Skin Cancer Screening Research Melanoma Treatment (PDQ®)–Patient Version General Information About Melanoma Key ... Certain factors affect prognosis (chance of recovery) and treatment options. The prognosis (chance of recovery ) and treatment ...

  9. Activation of endogenous p53 by combined p19Arf gene transfer and nutlin-3 drug treatment modalities in the murine cell lines B16 and C6

    International Nuclear Information System (INIS)

    Reactivation of p53 by either gene transfer or pharmacologic approaches may compensate for loss of p19Arf or excess mdm2 expression, common events in melanoma and glioma. In our previous work, we constructed the pCLPG retroviral vector where transgene expression is controlled by p53 through a p53-responsive promoter. The use of this vector to introduce p19Arf into tumor cells that harbor p53wt should yield viral expression of p19Arf which, in turn, would activate the endogenous p53 and result in enhanced vector expression and tumor suppression. Since nutlin-3 can activate p53 by blocking its interaction with mdm2, we explored the possibility that the combination of p19Arf gene transfer and nutlin-3 drug treatment may provide an additive benefit in stimulating p53 function. B16 (mouse melanoma) and C6 (rat glioma) cell lines, which harbor p53wt, were transduced with pCLPGp19 and these were additionally treated with nutlin-3 or the DNA damaging agent, doxorubicin. Viral expression was confirmed by Western, Northern and immunofluorescence assays. p53 function was assessed by reporter gene activity provided by a p53-responsive construct. Alterations in proliferation and viability were measured by colony formation, growth curve, cell cycle and MTT assays. In an animal model, B16 cells were treated with the pCLPGp19 virus and/or drugs before subcutaneous injection in C57BL/6 mice, observation of tumor progression and histopathologic analyses. Here we show that the functional activation of endogenous p53wt in B16 was particularly challenging, but accomplished when combined gene transfer and drug treatments were applied, resulting in increased transactivation by p53, marked cell cycle alteration and reduced viability in culture. In an animal model, B16 cells treated with both p19Arf and nutlin-3 yielded increased necrosis and decreased BrdU marking. In comparison, C6 cells were quite susceptible to either treatment, yet p53 was further activated by the combination of p19

  10. Notch ligand Delta-like 1 promotes the metastasis of melanoma by enhancing tumor adhesion

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, J.P. [Department of Orthopedic Surgery, Tangdu Hospital, The Fourth Military Medical University, Xi' an (China); Li, N. [Department of Oncology, Tangdu Hospital, The Fourth Military Medical University, Xi' an (China); Bai, W.Z.; Qiu, X.C.; Ma, B.A.; Zhou, Y.; Fan, Q.Y.; Shan, L.Q. [Department of Orthopedic Surgery, Tangdu Hospital, The Fourth Military Medical University, Xi' an (China)

    2014-03-28

    Notch signaling plays a vital role in tumorigenicity and tumor progression by regulating proliferation, invasion, and the tumor microenvironment. Previous research by our group indicated that Notch ligand Delta-like 1 (Dll1) is involved in angiogenesis in melanoma, and we noticed that it took a longer time to trypsinize Dll1-expressing B16 melanoma cells than the control cells. In this article, we extended our study to investigate the effects of Dll1 on tumor cell adhesion and metastasis. Dll1 overexpression activated Notch signaling in B16 tumor cells and significantly enhanced the adhering capacity of B16 tumor cells both in vitro and in vivo. B16-Dll1 cells also had a higher metastatic potential than their counterpart in the mouse model of lung metastasis. Along with increased Dll1 expression, N-cadherin, but not E-cadherin, was upregulated in B16-Dll1 cells. These data suggested that Notch ligand Dll1 may enhance the adhesion and metastasis of melanoma cells by upregulation of N-cadherin.

  11. Are all melanomas dangerous?

    DEFF Research Database (Denmark)

    Nørgaard, Carsten; Glud, Martin; Gniadecki, Robert

    2011-01-01

    The increased incidence of cutaneous malignant melanoma, together with only minor changes in mortality, has brought into question the existence of a melanoma epidemic. The discrepancy between incidence and mortality suggests that most newly diagnosed melanomas have indolent behaviour. This review...

  12. Genetics of familial melanoma

    DEFF Research Database (Denmark)

    Aoude, Lauren G; Wadt, Karin A W; Pritchard, Antonia L;

    2015-01-01

    Twenty years ago, the first familial melanoma susceptibility gene, CDKN2A, was identified. Two years later, another high-penetrance gene, CDK4, was found to be responsible for melanoma development in some families. Progress in identifying new familial melanoma genes was subsequently slow; however...

  13. Burden of Melanoma

    NARCIS (Netherlands)

    C. Holterhues (Cynthia)

    2011-01-01

    markdownabstract__Abstract__ Melanoma is a type of skin cancer that arises from melanocytes. More than 95% of all melanomas occur in the skin, but rarely in the pigmented cells of the eye, meninges or mucosa. This thesis will only regard the invasive cutaneous malignant melanomas.

  14. Radioiodide uptake in melanoma cells after transfer of human NaI symporter gene

    International Nuclear Information System (INIS)

    To obtain human sodium/iodide symporter gene cDNA for studying its potential ability as a radioiodide treatment for melanoma, the hNIS gene cDNA was amplified with total RNA from human thyroid tissue by RT-PCR. The hNIS cDNA was inserted into cloning vector pUCm-T and subcloned into eukaryotic expression vector pc-DNA3. The pc-DNA3-hNIS and pc-DNA3 were transduced into melanoma cells (B16) by electroporation, and two cell lines termed B16-A and B16-B respectively were established. The uptake and efflux of iodide was examined in vitro. The three cell lines (B16-A, B16-B, B16) were injected subcutaneously into the right flank of C57 mice. Biodistribution study and tumor imaging were performed when the tumor reached approximately 10 mm in diameter. The cloned hNIS cDNA sequence was identical with the published sequence. Two novel cell lines named B16-A containing pc-DNA3-hNIS and B16-B containing pc-DNA3 only were established. The resultant cell line B16-A accumulated 17 and 19 times more radioiodide in vitro than B16 and B16-B respectively. The iodide uptake reached the half-maximal level within 10 min, and reached a plateau at 30 min. The efflux of iodide was also rapid (T1/2eff=10 min). The imaging shows in vivo uptake in expected sites including the salivary glands, thyroid, stomach, and hNIS-transduced tumor, whereas the nontransduced tumor was not visualized. The %ID/g of B16-A tumors at 1,2,4,12 and 24h after injection of 125I were 12.22 ± 0.71, 10.91 ± 0.72, 8.73 ± 0.99, 1.24 ± 0.29, and 0.19 ± 0.03, respectively, which were significantly higher percentages than those for controlling tumors, p1/2 was about 6 h. Our preliminary data indicate that the transduction of the hNIS gene per se is sufficient to induce iodide transport in melanoma cells both in vitro and in vivo, but T1/2eff is short. (authors)

  15. Melanoma Cell Expression of CD200 Inhibits Tumor Formation and Lung Metastasis via Inhibition of Myeloid Cell Functions

    OpenAIRE

    Talebian, Fatemeh; Liu, Jin-Qing; Liu, Zhenzhen; Khattabi, Mazin; He, Yukai; Ganju, Ramesh; Bai, Xue-feng

    2012-01-01

    CD200 is a cell surface glycoprotein that functions through engaging CD200 receptor on cells of the myeloid lineage and inhibits their functions. Expression of CD200 has been implicated in a variety of human cancer cells including melanoma cells and has been thought to play a protumor role. To investigate the role of cancer cell expression of CD200 in tumor formation and metastasis, we generated CD200-positive and CD200-negative B16 melanoma cells. Subcutaneous injection of CD200-positive B16...

  16. Novel alpha-MSH peptide analogs for melanoma targeting

    Science.gov (United States)

    Flook, Adam Michael

    Skin cancer is the one of the most diagnosed cancers in the United States with increasing incidence over the past two decades. There are three major forms of skin cancer but melanoma is the deadliest. It is estimated that 76,690 new diagnoses of melanoma and 9,480 deaths will occur in 2013. Melanoma accounts for approximately 1.6% of all cancer related deaths and is the 5 th leading diagnosed cancer in the United States. The mean survival rate of patients diagnosed with metastatic melanoma is six months, with five year survival rates of less than 5%. In this project, we describe the design and characterization of novel melanoma-targeting peptide analogs for use in diagnostic imaging of both primary and metastatic melanoma lesions. Novel alpha-MSH peptide conjugates were designed to target the melanocortin-1 receptor present and over-expressed on melanoma cells. These peptides were synthesized and their in-vitro melanocortin-1 receptor binding affinities were established in murine melanoma cells. Once binding affinities were determined, the peptides were radiolabeled with 99mTc utilizing a novel direct radiolabeling technique developed in our laboratory. The peptides were purified via reverse-phase high performance liquid chromatography and in-vivo melanoma targeting and pharmacokinetic properties were determined in B16/F1 melanoma-bearing female C57BL/6 mice. Biodistribution and SPECT/CT imaging studies were performed with the promising 99m Tc-labeled peptide conjugates. All alpha-MSH peptide conjugates tested showed low nanomolar binding affinity for the melanocortin-1 receptor. All peptides were readily radiolabeld with 99mTc with greater than 95% radiochemical purity. All 99mTc-labeled peptides displayed high specific in-vivo melanoma tumor uptake while maintaining low normal organ accumulation, and were excreted through the urinary system in a timely fashion. In addition, all tested 99mTc-labeld alpha-MSH peptides demonstrated clear visualization of in

  17. Inhibitory Effect of Doxycycline on the Adhesion of Melanoma Cells and Its MolecularMechanisms%多西环素抑制黑色素瘤细胞黏附的分子机制研究

    Institute of Scientific and Technical Information of China (English)

    董恒磊; 孙保存; 孙涛; 赵楠; 董学易; 车娜; 赵秀兰

    2011-01-01

    Objective: To investigate the inhibitory effect of Doxycycline on the adhesion of melanoma cells and its mechanism.Methods: Melanoma B16F110, WM351 and WM451 cell lines were treated with Doxycycline at different doses and through different administration methods.MMT, cell adhesion experiment and Western blot were used to observe the effect of different treatment.A total of 20 C57/BL mice were transplanted with melanoma through injection of B16F10 cells and were divided into two groups.One group was treated with Doxycycline, and the other NaCl solution.Carcinoma tissues were obtained and underwentWestern blot and gelatin zymography.Results: MTT analysis showed that the inhibition ratio was significantly higher in the group receiving treatment before cell adhesion than in the group receiving treatment after cell adhesion ( P < 0.05 ).Cell adhesion experiment showed that Doxycycline had an inhibitory effect on the adhesion of B16F110, WM351 and WM451 cells, with a statistical significance ( P < 0.05 ).Western blot revealed that FAK expression in the three cell lines was decreased at 12 h after Doxycycline treatment.Observations from the mice melanoma model suggested obvious changes in FAK expression and its phosphorylation level ( P < 0.05 ).Conclusion: Doxycycline can interfere the adhesion of melanoma cells, possibly through inhibiting FAK expression and its phosphorylation.%目的:研究多西环素对黑色素瘤细胞黏附的抑制作用及分子机制.方法:对黑色素瘤细胞系B16F10、WM351、WM451分别以不同给药方式和给药浓度进行处理,利用MTT实验、细胞黏附实验和Western blot技术检测比较其差异.C57/BL小鼠20只进行黑色素瘤动物实验,接种B16F10后随机分为2组,分别给予多西环素和NaCl溶液处理,取瘤组织进行Western blot和明胶酶谱检测.结果:MTT实验显示"贴壁前"给药组的细胞抑制率显著高于"贴壁后"给药组且差异具有统计学意义(P<0.05).细胞黏附实

  18. Imaging malignant melanoma with {sup 18}F-5-FPN

    Energy Technology Data Exchange (ETDEWEB)

    Feng, Hongyan; Xia, Xiaotian; Li, Chongjiao; Song, Yiling; Qin, Chunxia; Liu, Qingyao; Zhang, Yongxue; Lan, Xiaoli [Department of Nuclear Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology (China); Hubei Key Laboratory of Molecular Imaging (China)

    2016-01-15

    Radiolabelled benzamides are attractive candidates for targeting melanoma because they bind to melanin and exhibit high tumour uptake and retention. {sup 18}F-5-Fluoro-N-(2-[diethylamino]ethyl)picolinamide ({sup 18}F-5-FPN), a benzamide analogue, was prepared and its pharmacokinetics and binding affinity evaluated both in vitro and in vivo to assess its clinical potential in the diagnosis and staging of melanoma. {sup 18}F-5-FPN was prepared and purified. Its binding specificity was measured in vitro in two different melanoma cell lines, one pigmented (B16F10 cells) and one nonpigmented (A375m cells), and in vivo in mice xenografted with the same cell lines. Dynamic and static PET images using {sup 18}F-5-FPN were obtained in the tumour-bearing mice, and the static images were also compared with those acquired with {sup 18}F-FDG. PET imaging with {sup 18}F-5-FPN was also performed in B16F10 tumour-bearing mice with lung metastases. {sup 18}F-5-FPN was successfully prepared with radiochemical yields of 5 - 10 %. Binding of {sup 18}F-5-FPN to B16F10 cells was much higher than to A375m cells. On dynamic PET imaging B16F10 tumours were visible about 1 min after injection of the tracer, and the uptake gradually increased over time. {sup 18}F-5-FPN was rapidly excreted via the kidneys. B16F10 tumours were clearly visible on static images acquired 1 and 2 h after injection, with high uptake values of 24.34 ± 6.32 %ID/g and 16.63 ± 5.41 %ID/g, respectively, in the biodistribution study (five mice). However, there was no visible uptake by A375m tumours. {sup 18}F-5-FPN and {sup 18}F-FDG PET imaging were compared in B16F10 tumour xenografts, and the tumour-to-background ratio of {sup 18}F-5-FPN was ten times higher than that of {sup 18}F-FDG (35.22 ± 7.02 vs. 3.29 ± 0.53, five mice). {sup 18}F-5-FPN PET imaging also detected simulated lung metastases measuring 1 - 2 mm. {sup 18}F-5-FPN specifically targeted melanin in vitro and in vivo with high retention and affinity

  19. Label-free detection of circulating melanoma cells by in vivo photoacoustic flow cytometry

    Science.gov (United States)

    Wang, Xiaoling; Yang, Ping; Liu, Rongrong; Niu, Zhenyu; Suo, Yuanzhen; He, Hao; Gao, Wenyuan; Tang, Shuo; Wei, Xunbin

    2016-03-01

    Melanoma is a malignant tumor of melanocytes. Melanoma cells have high light absorption due to melanin highly contained in melanoma cells. This property is employed for the detection of circulating melanoma cell by in vivo photoacoustic flow cytometry (PAFC), which is based on photoacoustic effect. Compared to in vivo flow cytometry based on fluorescence, PAFC can employ high melanin content of melanoma cells as endogenous biomarkers to detect circulating melanoma cells in vivo. We have developed in vitro experiments to prove the ability of PAFC system of detecting photoacoustic signals from melanoma cells. For in vivo experiments, we have constructed a model of melanoma tumor bearing mice by inoculating highly metastatic murine melanoma cancer cells, B16F10 with subcutaneous injection. PA signals are detected in the blood vessels of mouse ears in vivo. The raw signal detected from target cells often contains some noise caused by electronic devices, such as background noise and thermal noise. We choose the Wavelet denoising method to effectively distinguish the target signal from background noise. Processing in time domain and frequency domain would be combined to analyze the signal after denoising. This algorithm contains time domain filter and frequency transformation. The frequency spectrum image of the signal contains distinctive features that can be used to analyze the property of target cells or particles. The processing methods have a great potential for analyzing signals accurately and rapidly. By counting circulating melanoma cells termly, we obtain the number variation of circulating melanoma cells as melanoma metastasized. Those results show that PAFC is a noninvasive and label-free method to detect melanoma metastases in blood or lymph circulation.

  20. Inhibition of melanocortin 1 receptor slows melanoma growth, reduces tumor heterogeneity and increases survival.

    Science.gov (United States)

    Kansal, Rita G; McCravy, Matthew S; Basham, Jacob H; Earl, Joshua A; McMurray, Stacy L; Starner, Chelsey J; Whitt, Michael A; Albritton, Lorraine M

    2016-05-01

    Melanoma risk is increased in patients with mutations of melanocortin 1 receptor (MC1R) yet the basis for the increased risk remains unknown. Here we report in vivo evidence supporting a critical role for MC1R in regulating melanoma tumor growth and determining overall survival time. Inhibition of MC1R by its physiologically relevant competitive inhibitor, agouti signaling protein (ASIP), reduced melanin synthesis and morphological heterogeneity in murine B16-F10 melanoma cells. In the lungs of syngeneic C57BL/6 mice, mCherry-marked, ASIP-secreting lung tumors inhibited MC1R on neighboring tumors lacking ASIP in a dose dependent manner as evidenced by a proportional loss of pigment in tumors from mice injected with 1:1, 3:1 and 4:1 mixtures of parental B16-F10 to ASIP-expressing tumor cells. ASIP-expressing B16-F10 cells formed poorly pigmented tumors in vivo that correlated with a 20% longer median survival than those bearing parental B16-F10 tumors (p=0.0005). Mice injected with 1:1 mixtures also showed survival benefit (p=0.0054), whereas injection of a 4:1 mixture showed no significant difference in survival. The longer survival time of mice bearing ASIP-expressing tumors correlated with a significantly slower growth rate than parental B16-F10 tumors as judged by quantification of numbers of tumors and total tumor load (p=0.0325), as well as a more homogeneous size and morphology of ASIP-expressing lung tumors. We conclude that MC1R plays an important role in regulating melanoma growth and morphology. Persistent inhibition of MC1R provided a significant survival advantage resulting in part from slower tumor growth, establishing MC1R as a compelling new molecular target for metastatic melanoma. PMID:27028866

  1. The leaf extract of Mallotus japonicus and its major active constituent, rutin,suppressed on melanin production in murine B16F1 melanoma简

    Institute of Scientific and Technical Information of China (English)

    Junsei; Taira; Eito; Tsuchida; Masatsugu; Uehara; Natsuko; Ohhama; Wakana; Ohmine; Takayuki; Ogi

    2015-01-01

    Objective: To find anti-melanogenesis materials used in whitening cosmetics.Methods: The ethanolic leaf extract of Mallotus japonicus(M. japonicus) having an anti-melanogenesis activity was separated by a sephadex LH-20 chromatography. Each fraction was measured for its tyrosinase inhibitory activity together with its polyphenol content using the Folin–Ciocalteu method. The anti-melanogenesis activity of the active fractions was determined by the melanin content in the murine B16F1 melanoma. The active fractions were put together due to similar constituents, and then separated by high performance liquid chromatography using a C-18 ODS column. The major antimelanogenesis compound was identified using1 H and13C-NMR and liquid chromatography-mass spectrometry.Results: The ethanolic leaf extract of M. japonicus showed an anti-tyrosinase activity with a high polyphenol content, resulting in suppression of melanin production in the B16F1 melanoma. The extract was separated and the active compound was identical as rutin based on the1 H,13C-NMR and liquid chromatography-mass spectrometry analysis data. In addition, the rutin treatment with cells reduced the melanin content in a concentration dependent manner without any cell toxicity. The leaf extract of M. japonicus containing rutin would be useful in whitening cosmetics for protection from UV-light exposure to be limiting the accumulation of melanin in skin.Conclusions: The leaf extract of M. japonicus and/or rutin isolated from the extract as a key whitening agent would be useful as a whitening cosmetic material for protecting against disorder skin due to melanin accumulation.

  2. Estudio tribológico comparativo de los Babbits Fórmula V y B-16 // Babbits comparative Study About Fórmulas V and B-16

    Directory of Open Access Journals (Sweden)

    M. Díaz Díaz

    1999-01-01

    Full Text Available En el trabajo se realiza un estudio comparativo del coeficiente de fricción de los Babbits, fórmula V y B-16, así como lainfluencia que ejerce sobre estos la presión de trabajo y el ángulo de contacto entre el cojinete y el árbol, determinándose lasexpresiones empíricas que rigen esta relación para ambos materiales antifricción.Palabras claves: Babbi t s, presión, angulo de contacto, coj inetes de desl izamiento._________________________________________________________________________________AbstractIn this work is made a comparative study about the friction coefficients of Babbits metals, Fórmula V and B-16, taking intoaccost influence the pressure and contact angle between shaft and bearing. Empirical relations for both antifriction materials aregiven.Key words: Babbi t s, pressure, contact angle, plain bearings

  3. Treatment of mouse melanoma cells with phorbol 12-myristate 13-acetate counteracts mannosylerythritol lipid-induced growth arrest and apoptosis

    OpenAIRE

    Zhao, Xiaoxian; Geltinger, Christian; Kishikawa, Shotaro; Ohshima, Kiyoshi; Murata, Takehide; Nomura, Nobuhiko; Nakahara, Tadaatsu; Yokoyama, Kazunari K.

    2000-01-01

    Mannosylerythritol lipid (MEL), an extracellularglycolipid from yeast, induces the differentiation ofHL-60 promyelocytic leukemia cells towardsgranulocytes. We show here that MEL is also a potentinhibitor of the proliferation of mouse melanoma B16cells. Flow-cytometric analysis of the cell cycle ofMEL-treated B16 cells revealed the accumulation ofcells in the sub-G0/G1 phase, which is a hallmark ofcells undergoing apoptosis. Treatment of B16 cellsfor 24 h with phorbol 12-myristate 13-acetate ...

  4. Nanodiamonds coupled with 5,7-dimethoxycoumarin, a plant bioactive metabolite, interfere with the mitotic process in B16F10 cells altering the actin organization

    Directory of Open Access Journals (Sweden)

    Gismondi A

    2016-02-01

    Full Text Available Angelo Gismondi,1 Valentina Nanni,1 Giacomo Reina,2 Silvia Orlanducci,2 Maria Letizia Terranova,2 Antonella Canini1 1Department of Biology, 2Department of Chemical Science and Technology, University of Rome “Tor Vergata”, Rome, Italy Abstract: For the first time, we coupled reduced detonation nanodiamonds (NDs with a plant secondary metabolite, citropten (5,7-dimethoxycoumarin, and demonstrated how this complex was able to reduce B16F10 tumor cell growth more effectively than treatment with the pure molecule. These results encouraged us to find out the specific mechanism underlying this phenomenon. Internalization kinetics and quantification of citropten in cells after treatment with its pure or ND-conjugated form were measured, and it was revealed that the coupling between NDs and citropten was essential for the biological properties of the complex. We showed that the adduct was not able to induce apoptosis, senescence, or differentiation, but it determined cell cycle arrest, morphological changes, and alteration of mRNA levels of the cytoskeletal-related genes. The identification of metaphasic nuclei and irregular disposition of β-actin in the cell cytoplasm supported the hypothesis that citropten conjugated with NDs showed antimitotic properties in B16F10 cells. This work can be considered a pioneering piece of research that could promote and support the biomedical use of plant drug-functionalized NDs in cancer therapy. Keywords: citropten, cytoskeletal structure, plant secondary metabolite, melanoma, internalization kinetics

  5. Radioiodide uptake in melanoma cells after transfer of human NaI symporter gene

    Institute of Scientific and Technical Information of China (English)

    CHEN Li-Bo; ZHU Rui-Sen; LU Han-Kui; YU Yong-Li; LUO Quan-Yong; HUANG Fang; FEI Jian; GUO Li-He

    2004-01-01

    To obtain human sodium/iodide symporter gene cDNA for studying its potential ability as a radioiodide treatment for melanoma, the hNIS gene cDNA was amplified with total RNA from human thyroid tissue by RT-PCR.The hNIS cDNA was inserted into cloning vector pUCm-T and subcloned into eukaryotic expression vector pc-DNA3.The pc-DNA3-hNIS and pc-DNA3 were transduced into melanoma cells (B 16) by electroporation, and two cell lines termed B 16-A and B16-B respectively were established. The uptake and efflux of iodide was examined in vitro. The three cell lines (B16-A, B16-B, B16) were injected subcutaneously into the right flank of C57 mice. Biodistribution study and tumor imaging were performed when the tumor reached approximately 10mm in diameter. The cloned hNIS cDNA sequence was identical with the published sequence. Two novel cell lines named 16-A containing pc-DNA3-hNIS and B16-B containing pc-DNA3 only were established. The resultant cell line B16-A accumulated 17and 19 times more radioiodide in vitro than B 16 and B 16-B respectively. The iodide uptake reached the half-maximal level within 10 min, and reached a plateau at 30 min. The efflux of iodide was also rapid (T1/2cff=10min). The imaging shows in vivo uptake in expected sites including the salivary glands, thyroid, stomach, and hNIS-transduced tumor,whereas the nontransduced tumor was not visualized. The %ID/g of B 16-A tumors at 1, 2, 4, 12, and 24h after injection of 125I were 12.22±0.71, 10.91±0.72, 8.73±0.99, 1.24±0.29, and 0.19±0.03, respectively, which were signifi cantly higher percentages than those for controlling tumors, p<0.01. However, biologic T1/2 was about 6 h. Our preliminary data indicate that the transduction of the hNIS gene per se is sufficient to induce iodide transport in melanoma cells both in vitro and in vivo, but T1/2eff is short.

  6. Effect of homeopathic medicines on transplanted tumors in mice.

    Science.gov (United States)

    Es, Sunila; Kuttan, Girija; Kc, Preethi; Kuttan, Ramadasan

    2007-01-01

    Ultra low doses used in homeopathic medicines are reported to have healing potential for various diseases but their action remains controversial. In this study we have investigated the antitumour and antimetastatic activity of selected homeopathic medicines against transplanted tumours in mice. It was found that Ruta graveolens 200c and Hydrastis canadensis 200c significantly increased the lifespan of Ehrlich Ascites Carcinoma and Dalton's Lymphoma Ascites induced tumour-bearing animals by 49.7%, and 69.4% respectively. Moreover there was 95.6% and 95.8% reduction of solid tumour volume in Ruta 200c and Hydrastis 200c treated animals on the 31st day after tumour inoculation. Hydrastis 1M given orally significantly inhibited the growth of developed solid tumours produced by DLA cells and increased the lifespan of tumour bearing animals. Some 9 out of 15 animals with developed tumors were completely tumour free after treatment with Hydrastis 1M. Significant anti-metastatic activity was also found in B16F-10 melanoma-bearing animals treated with Thuja1M, Hydrastis 1M and Lycopodium1M. This was evident from the inhibition of lung tumour nodule formation, morphological and histopathological analysis of lung and decreased levels of gamma-GT in serum, a cellular marker of proliferation. These findings support that homeopathic preparations of Ruta and Hydrastis have significant antitumour activity. The mechanism of action of these medicines is not known at present. PMID:18159975

  7. Evaluation of two (125)I-radiolabeled acridine derivatives for Auger-electron radionuclide therapy of melanoma.

    Science.gov (United States)

    Gardette, Maryline; Viallard, Claire; Paillas, Salomé; Guerquin-Kern, Jean-Luc; Papon, Janine; Moins, Nicole; Labarre, Pierre; Desbois, Nicolas; Wong-Wah-Chung, Pascal; Palle, Sabine; Wu, Ting-Di; Pouget, Jean-Pierre; Miot-Noirault, Elisabeth; Chezal, Jean-Michel; Degoul, Francoise

    2014-08-01

    We previously selected two melanin-targeting radioligands [(125)I]ICF01035 and [(125)I]ICF01040 for melanoma-targeted (125)I radionuclide therapy according to their pharmacological profile in mice bearing B16F0 tumors. Here we demonstrate in vitro that these compounds present different radiotoxicities in relation to melanin and acidic vesicle contents in B16F0, B16F0 PTU and A375 cell lines. ICF01035 is effectively observed in nuclei of achromic (A375) melanoma or in melanosomes of melanized melanoma (B16F0), while ICF01040 stays in cytoplasmic vesicles in both cells. [(125)I]ICF01035 induced a similar survival fraction (A50) in all cell lines and led to a significant decrease in S-phase cells in amelanotic cell lines. [(125)I]ICF01040 induced a higher A50 in B16 cell lines compared to [(125)I]ICF01035 ones. [(125)I]ICF01040 induced a G2/M blockade in both A375 and B16F0 PTU, associated with its presence in cytoplasmic acidic vesicles. These results suggest that the radiotoxicity of [(125)I]ICF01035 and [(125)I]ICF01040 are not exclusively reliant on DNA alterations compatible with γ rays but likely result from local dose deposition (Auger electrons) leading to toxic compound leaks from acidic vesicles. In vivo, [(125)I]ICF01035 significantly reduced the number of B16F0 lung colonies, enabling a significant increase in survival of the treated mice. Targeting melanosomes or acidic vesicles is thus an option for future melanoma therapy. PMID:24691673

  8. Stress indices for ANSI standard B16.11 socket-welding fittings

    International Nuclear Information System (INIS)

    Stress indices for ANSI standard B16.11 socket-welding tees, 450 elbows, 900 elbows, and couplings are developed for intended use with the Class-1 piping system design rules of Section III--Division 1 of the ASME Boiler and Pressure Vessel Code. Indices are given for the evaluation of appropriate primary stresses, primary-plus-secondary stresses, and peak stresses due to internal pressure, bending-moment loads, and thermal gradients between the fitting and the attached pipe. The proposed indices are based on the dimensional and pressure-burst requirements of the B16.11 standard, the apparent shapes of B16.11 fittings as indicated from a random sampling taken off-the-shelf, the standard pressure-temperature ratings of the fittings, and on current stress indices now in the Code for similar butt-welding fittings. Specific recommendations are made for issuing the new stress indices in a Code case. (auth)

  9. Decoding Melanoma Metastasis

    Energy Technology Data Exchange (ETDEWEB)

    Damsky, William E. Jr. [Department of Dermatology, Yale School of Medicine, New Haven, Connecticut (United States); Department of Pathology, University of Vermont College of Medicine, Burlington, Vermont (United States); Rosenbaum, Lara E.; Bosenberg, Marcus, E-mail: Marcus.Bosenberg@yale.edu [Department of Dermatology, Yale School of Medicine, New Haven, Connecticut (United States)

    2010-12-30

    Metastasis accounts for the vast majority of morbidity and mortality associated with melanoma. Evidence suggests melanoma has a predilection for metastasis to particular organs. Experimental analyses have begun to shed light on the mechanisms regulating melanoma metastasis and organ specificity, but these analyses are complicated by observations of metastatic dormancy and dissemination of melanocytes that are not yet fully malignant. Additionally, tumor extrinsic factors in the microenvironment, both at the site of the primary tumor and the site of metastasis, play important roles in mediating the metastatic process. As metastasis research moves forward, paradigms explaining melanoma metastasis as a step-wise process must also reflect the temporal complexity and heterogeneity in progression of this disease. Genetic drivers of melanoma as well as extrinsic regulators of disease spread, particularly those that mediate metastasis to specific organs, must also be incorporated into newer models of melanoma metastasis.

  10. Estudio tribológico comparativo de los Babbits Fórmula V y B-16 // Babbits comparative Study About Fórmulas V and B-16

    OpenAIRE

    M. Díaz Díaz; C. Díaz Novo; O. Calderín Medina

    1999-01-01

    En el trabajo se realiza un estudio comparativo del coeficiente de fricción de los Babbits, fórmula V y B-16, así como lainfluencia que ejerce sobre estos la presión de trabajo y el ángulo de contacto entre el cojinete y el árbol, determinándose lasexpresiones empíricas que rigen esta relación para ambos materiales antifricción.Palabras claves: Babbi t s, presión, angulo de contacto, coj inetes de desl izamiento._________________________________________________________________________________...

  11. Anti-tumor Properties of cis-Resveratrol Methylated Analogues in Metastatic Mouse Melanoma Cells

    Science.gov (United States)

    Morris, Valery L.; Toseef, Tayyaba; Nazumudeen, Fathima B.; Rivoira, Christian; Spatafora, Carmela; Tringali, Corrado; Rotenberg, Susan A.

    2015-01-01

    Resveratrol (E-3,5,4’-trihydroxystilbene) is a polyphenol found in red wine that has been shown to have multiple anti-cancer properties. Although cis (Z) and trans (E) isomers of resveratrol occur in nature, the cis form is not biologically active. However, methylation at key positions of the cis form results in more potent anti-cancer properties. This study determined that synthetic cis-polymethoxystilbenes (methylated analogues of cis-resveratrol) inhibited cancer-related phenotypes of metastatic B16 F10 and non-metastatic B16 F1 mouse melanoma cells. In contrast with cis or trans-resveratrol and trans-polymethoxystilbene which were ineffective at 10 μM, cis-polymethoxystilbenes inhibited motility and proliferation of melanoma cells with low micromolar specificity (IC50 <10 μM). Inhibitory effects by cis-polymethoxystilbenes were significantly stronger with B16 F10 cells and were accompanied by decreased expression of β-tubulin and pleckstrin homology domain-interacting protein, a marker of metastatic B16 cells. Thus, cis-polymethoxystilbenes have potential as chemotherapeutic agents for metastatic melanoma. PMID:25567208

  12. Effect of Adipocyte Secretome in Melanoma Progression and Vasculogenic Mimicry.

    Science.gov (United States)

    Coelho, Pedro; Almeida, Joana; Prudêncio, Cristina; Fernandes, Rúben; Soares, Raquel

    2016-07-01

    Obesity, favored by the modern lifestyle, acquired epidemic proportions nowadays. Obesity has been associated with various major causes of death and morbidity including malignant neoplasms. This increased prevalence has been accompanied by a worldwide increase in cutaneous melanoma incidence rates during the last decades. Obesity involvement in melanoma aetiology has been recognized, but the implicated mechanisms remain unclear. In the present study, we address this relationship and investigate the influence of adipocytes secretome on B16-F10 and MeWo melanoma cell lines. Using the 3T3-L1 adipocyte cell line, as well as ex vivo subcutaneous (SAT) and visceral (VAT) adipose tissue conditioned medium, we were able to show that adipocyte-released factors play a dual role in increasing melanoma cell overall survival, both by enhancing proliferation and decreasing apoptosis. B16-F10 cell migration and cell-cell and cell-matrix adhesion capacity were predominantly enhanced in the presence of SAT and VAT released factors. Melanocytes morphology and melanin content were also altered by exposure to adipocyte conditioned medium disclosing a more dedifferentiated phenotype of melanocytes. In addition, exposure to adipocyte-secreted molecules induced melanocytes to rearrange, on 3D cultures, into vessel-like structures, and generate characteristic vasculogenic mimicry patterns. These findings are corroborated by the released factors profile of 3T3-L1, SAT, and VAT assessed by microarrays, and led us to highlight the mechanisms by which adipose secretome from sub-cutaneous or visceral depots promote melanoma progression. J. Cell. Biochem. 117: 1697-1706, 2016. © 2015 Wiley Periodicals, Inc. PMID:26666522

  13. Ctla-4 blockade plus adoptive T-cell transfer promotes optimal melanoma immunity in mice.

    Science.gov (United States)

    Mahvi, David A; Meyers, Justin V; Tatar, Andrew J; Contreras, Amanda; Suresh, Marulasiddappa; Leverson, Glen E; Sen, Siddhartha; Cho, Clifford S

    2015-01-01

    Immunotherapeutic approaches to the treatment of advanced melanoma have relied on strategies that augment the responsiveness of endogenous tumor-specific T-cell populations [eg, cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) blockade-mediated checkpoint inhibition] or introduce exogenously prepared tumor-specific T-cell populations [eg, adoptive cell transfer (ACT)]. Although both approaches have shown considerable promise, response rates to these therapies remain suboptimal. We hypothesized that a combinatorial approach to immunotherapy using both CTLA-4 blockade and nonlymphodepletional ACT could offer additive therapeutic benefit. C57BL/6 mice were inoculated with syngeneic B16F10 melanoma tumors transfected to express low levels of the lymphocytic choriomeningitis virus peptide GP33 (B16GP33), and treated with no immunotherapy, CTLA-4 blockade, ACT, or combination immunotherapy of CTLA-4 blockade with ACT. Combination immunotherapy resulted in optimal control of B16GP33 melanoma tumors. Combination immunotherapy promoted a stronger local immune response reflected by enhanced tumor-infiltrating lymphocyte populations, and a stronger systemic immune responses reflected by more potent tumor antigen-specific T-cell activity in splenocytes. In addition, whereas both CTLA-4 blockade and combination immunotherapy were able to promote long-term immunity against B16GP33 tumors, only combination immunotherapy was capable of promoting immunity against parental B16F10 tumors as well. Our findings suggest that a combinatorial approach using CTLA-4 blockade with nonlymphodepletional ACT may promote additive endogenous and exogenous T-cell activities that enable greater therapeutic efficacy in the treatment of melanoma.

  14. Bone marrow transplant

    Science.gov (United States)

    Transplant - bone marrow; Stem cell transplant; Hematopoietic stem cell transplant; Reduced intensity nonmyeloablative transplant; Mini transplant; Allogenic bone marrow transplant; Autologous bone marrow transplant; Umbilical ...

  15. Anti-tumor activity of N-trimethyl chitosan-encapsulated camptothecin in a mouse melanoma model

    Directory of Open Access Journals (Sweden)

    Yi Tao

    2010-06-01

    Full Text Available Abstract Background Camptothecin (CPT has recently attracted increasing attention as a promising anticancer agent for a variety of tumors. But the clinical application is largely hampered by its extreme water insolubility and unpredictable side effect. It is essential to establish an efficient and safe protocol for the administration of CPT versus melanoma. Methods Camptothecin was encapsulated with N-trimethyl chitosan (CPT-TMC through microprecipitation and sonication. Its inhibition effect on B16-F10 cell proliferation and induction of apoptosis was evaluated by MTT assay and flow cytometric analysis in vitro. The anti-tumor activity of CPT-TMC was evaluated in C57BL/6 mice bearing B16-F10 melanoma. Tumor volume, tumor weight and survival time were recorded. Assessment of apoptotic cells within tumor tissue was performed by TUNEL assay. Antiangiogenesis and antiproliferation effects of CPT-TMC in vivo were conducted via CD31 and PCNA immunohistochemistry, respectively. Results CPT-TMC efficiently inhibited B16-F10 cells proliferation and increased apoptosis in vitro. Experiment group showed significant inhibition compared with free CPT-treated group (81.3% vs. 56.9% in the growth of B16-F10 melanoma xenografts and prolonged the survival time of the treated mice (P Conclusions Our data suggest that N-trimethyl chitosan-encapsulated camptothecin is superior to free CPT by overcoming its insolubility and finally raises the potential of its application in melanoma therapy.

  16. Effects of hCD80-Adenovirus on B16-F10 Cells

    Institute of Scientific and Technical Information of China (English)

    田长富; 李殿俊; 刘旭

    2002-01-01

    @@ To study the biological characteristics of B16-F10 cells transduced with hCD80 gene in vitro, a kind of replication-deficient recombinant adenovirus bearing hCD80 gene (hCD80-rAd) was constructed and several means were utilized to observe the changes of biological characteristics after gene transduced.

  17. Comparative study of angio genesis radiopharmaceuticals for melanoma detection

    International Nuclear Information System (INIS)

    Early diagnosis and treatment of melanoma, a cutaneous tumor with a serious prognosis, is extremely important for optimal clinical outcome. Phage display peptide libraries are a useful screening resource for identifying bioactive peptides that interact with cancer targets. The aim of this study was the evaluation of two technetium-99m tracers for angio genesis detection in melanoma model, using cyclic peguilated pentapeptide with RGD and NGR motifs conjugated with bifunctional chelator MAG3. The conjugated peptides (10 μL of a μg/μL solution) were labeled with technetium-99m using a sodium tartrate buffer. Radiochemical evaluation was done by ITLC and confirmed by HPLC. Partition coefficient was determined and internalization assays were performed in two melanoma cells (B16F10 and SKMEL28). Biodistribution evaluation of the tracers was done in healthy animals at different times and also in mice bearing the tumor cells at 120 min post injection. Blocking studies were also conducted by co-injection of cold peptides. The conjugated showed the same profile in many evaluations. They were radiolabeled with high radiochemical purity (>97%). Both were hydrophilic, with preferential renal excretion. Tumor uptake was higher for human melanoma cells than for murinic melanoma cells, specially for 99mTc-MAG3-PEG8-c(RGDyK) (7.85±±2.34 %ID/g) at 120 min post injection. The performance of 99mTc-MAG3-PEG8-c(RGDyk) was much better than NGR tracer concerning human melanoma uptake and might be considered in future investigations focusing radiotracers for melanoma diagnosis. (author)

  18. Melanoma Therapy via Peptide-Targeted a-Radiation

    Energy Technology Data Exchange (ETDEWEB)

    Miao, Yubin; Hylarides, Mark; Fisher, Darrell R.; Shelton, Tiffani; Moore, Herbert A.; Wester, Dennis W.; Fritzberg, Alan R.; Winkelmann, Christopher T.; Hoffman, Timothy J.; Quinn, Thomas P.

    2005-08-01

    Malignant melanoma is the most lethal form of skin cancer. Current chemotherapy and external beam radiation therapy regimens are ineffective agents against melanoma, as shown by a 10-year survival rate for patients with disseminated disease of approximately 5% (reference?). In this study, the unique combination of a melanoma targeting peptide and an in vivo generated a-particle emitting radioisotope was investigated for its melanoma therapy potential. Alpha-radiation is densely ionizing and energy is locally absorbed, resulting in high concentrations of destructive free radicals and irreparable DNA double strand breaks. This high linear-energy-transfer overcomes radiation resistant tumor cells and oxygen-enhancement effects. The melanoma targeting peptide DOTA-Re(Arg11)CCMSH was radiolabeled with 212Pb, the parent of 212Bi, which decays via alpha and beta decay. Biodistribution and therapy studies were performed in the B16/F1 melanoma bearing C57 mouse flank tumor model. 212Pb[DOTA]-R e(Arg11)CCMSH exhibited rapid tumor uptake and extended retention coupled with rapid whole body disappearance. Radiation dose delivered to the tumor was estimated to be 61 cGy/uCi 212Pb administered. Treatment of melanoma-bearing mice with 50, 100 and 200 uCi of 212Pb[DOTA]-Re(Arg11)CCMSH extended mean survival of mice to 22, 28, and 49.8 days, respectively, compared to the 14.6 day mean survival of the placebo control group. Forty-five percent of the mice receiving 200 uCi survived the study disease-free.

  19. Effects of Genistein on Cell Cycle and Apoptosis of Two Murine Melanoma Cell Lines

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The effects of genistein on several tumor cell lines were investigated to study the effects of genistein on cell growth, cell cycle, and apoptosis of two murine melanoma cell lines, B16 and K1735M2. These two closely related murine melanoma cell lines, however, have different responses to the genistein treatment. Genistein inhibits the growth of both the B16 and K1735M2 cell lines and arrests the growth at the G2/M phase. After treatment with 60 μmol/L genistein for 72 h, apoptosis and caspase activities were detected in B16 cells, while such effects were not found in K1735M2. Further tests showed that after genistein treatment the protein content and mRNA levels of p53 increased in B16, but remained the same in K1735M2. The protein content and mRNA levels of p21WAF1/CIP1 increased in both cell lines after treatment.The results show that genistein might induce apoptosis in B16 cells by damaging the DNA, inhibiting topoisomerase Ⅱ, increasing p53 expression, releasing cytochrome c from the mitochondria, and activating the caspases which will lead to apoptosis.

  20. Adenoviral vaccination combined with CD40 stimulation and CTLA-4 blockage can lead to complete tumor regression in a murine melanoma model

    DEFF Research Database (Denmark)

    Sørensen, Maria Rathmann; Holst, Peter J; Steffensen, Maria Abildgaard;

    2010-01-01

    Therapeutic vaccination with replication deficient adenovirus expressing a viral antigen linked to invariant chain was recently found to markedly delay the growth of B16.F10 melanomas expressing the same antigen; however, complete regression of the tumors was never observed. Here we show that the......Therapeutic vaccination with replication deficient adenovirus expressing a viral antigen linked to invariant chain was recently found to markedly delay the growth of B16.F10 melanomas expressing the same antigen; however, complete regression of the tumors was never observed. Here we show...

  1. Translational research in melanoma.

    Science.gov (United States)

    Ray, Madhury; Farma, Jeffrey M; Hsu, Cary

    2013-10-01

    Recent breakthroughs in the fundamental understanding of the cellular and molecular basis of melanoma have culminated in new therapies with unquestionable efficacy. Immunotherapy and targeted therapy strategies have completely transformed the contemporary management of advanced melanoma. The translational research behind these developments is discussed, with an emphasis on immune checkpoint blockade and inhibition of the mitogen-activated protein kinase signaling pathway.

  2. Are all melanomas dangerous?

    DEFF Research Database (Denmark)

    Nørgaard, Carsten; Glud, Martin; Gniadecki, Robert

    2011-01-01

    incidence of melanoma during the last few decades, with by far the highest increase in tumours at a very early stage (T1 or IA). Little or no change was seen in mortality. However, increases in both mortality and incidence of thick melanomas were found in the oldest subgroups, especially in men. These...

  3. Docosahexaenoic acid inhibits melanin synthesis in murine melanoma cells in vitro through increasing tyrosinase degradation

    OpenAIRE

    Balcos, Marie Carmel; Kim, Su Yeon; Jeong, Hyo-Soon; Yun, Hye-Young; Baek, Kwang Jin; Kwon, Nyoun Soo; Park, Kyoung-Chan; Kim, Dong-Seok

    2014-01-01

    Aim: To investigate the effects of docosahexaenoic acid (DHA) on melanin synthesis and related regulatory mechanisms. Methods: B16F10 mouse melanoma cells were exposed to DHA for 3 d, and melanin content and tyrosinase activity were measured. Western blot analysis was used to analyze the protein levels in DHA-mediated signal transduction pathways. Results: DHA (1–25 μmol/L) did not affect the viability of B16F10 cells, but decreased α-MSH-induced melanin synthesis in a concentration-dependent...

  4. Familial malignant melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Kopf, A.W.; Hellman, L.J.; Rogers, G.S.; Gross, D.F.; Rigel, D.S.; Friedman, R.J.; Levenstein, M.; Brown, J.; Golomb, F.M.; Roses, D.F.; Gumport, S.L.

    1986-10-10

    Characteristics associated with familial compared with nonfamilial malignant melanoma were assessed. These data were obtained from consecutive prospectively completed questionnaires on 1169 cases of cutaneous malignant melanoma. Of these, 69 patients indicated a positive family history for this cancer. Among the various clinical and histological variables compared, those that significantly correlated with the familial occurrence of malignant melanoma include younger age at first diagnosis, smaller diameter of the lesion, lower Clark level, decreased frequency of nonmelanoma skin cancer, and reduced prevalence of noncutaneous cancer. Increased awareness of malignant melanoma among family members could account for some of these observations. Identification of the familial variety of malignant melanoma has practical implications concerning early detection and prompt intervention.

  5. Main roads to melanoma

    Directory of Open Access Journals (Sweden)

    Sini Maria

    2009-10-01

    Full Text Available Abstract The characterization of the molecular mechanisms involved in development and progression of melanoma could be helpful to identify the molecular profiles underlying aggressiveness, clinical behavior, and response to therapy as well as to better classify the subsets of melanoma patients with different prognosis and/or clinical outcome. Actually, some aspects regarding the main molecular changes responsible for the onset as well as the progression of melanoma toward a more aggressive phenotype have been described. Genes and molecules which control either cell proliferation, apoptosis, or cell senescence have been implicated. Here we provided an overview of the main molecular changes underlying the pathogenesis of melanoma. All evidence clearly indicates the existence of a complex molecular machinery that provides checks and balances in normal melanocytes. Progression from normal melanocytes to malignant metastatic cells in melanoma patients is the result of a combination of down- or up-regulation of various effectors acting on different molecular pathways.

  6. Expression and functions of galectin-7 in human and murine melanomas.

    Directory of Open Access Journals (Sweden)

    Katherine Biron-Pain

    Full Text Available The identification of galectin-7 as a p53-induced gene and its ability to induce apoptosis in many cell types support the hypothesis that galectin-7 has strong antitumor activity. This has been well documented in colon cancer. However, in some cases, such as breast cancer and lymphoma, its high expression level correlates with aggressive subtypes of cancer, suggesting that galectin-7 may have a dual role in cancer progression. In fact, in breast cancer, overexpression of galectin-7 alone is sufficient to promote metastasis to the bone and lung. In the present work, we investigated the expression and function of galectin-7 in melanoma. An analysis of datasets obtained from whole-genome profiling of human melanoma tissues revealed that galectin-7 mRNA was detected in more than 90% of biopsies of patients with nevi while its expression was more rarely found in biopsies collected from patients with malignant melanoma. This frequency, however, was likely due to the presence of normal epidermis tissues in biopsies, as shown our studies at the protein level by immunohistochemical analysis. Using the experimental melanoma B16F1 cell line, we found that melanoma cells can express galectin-7 at the primary tumor site and in lung metastasis. Moreover, we found that overexpression of galectin-7 increased the resistance of melanoma cells to apoptosis while inducing de novo egr-1 expression. Overexpression of galectin-7, however, was insufficient to modulate the growth of tumors induced by the subcutaneous injection of B16F1 cells. It also failed to modulate the dissemination of B16F1 cells to the lung.

  7. Liver Transplant

    Science.gov (United States)

    ... Home > Your Liver > Liver Disease Information > Liver Transplant Liver Transplant Explore this section to learn more about liver ... harmful substances from your blood. What is a liver transplant? A liver transplant is the process of replacing ...

  8. Synthesis and evaluation of novel radioiodinated nicotinamides for malignant melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Liu Xiang; Pham, Tien Q.; Berghofer, Paula; Chapman, Janette; Greguric, Ivan; Mitchell, Peter; Mattner, Filomena; Loc' h, Christian [Radiopharmaceuticals Research Institute, Australian Nuclear Science and Technology Organisation, Sydney, NSW 2234 (Australia); Katsifis, Andrew [Radiopharmaceuticals Research Institute, Australian Nuclear Science and Technology Organisation, Sydney, NSW 2234 (Australia)], E-mail: akx@ansto.gov.au

    2008-10-15

    Introduction: A series of iodonicotinamides based on the melanin-binding iodobenzamide compound N-2-diethylaminoethyl-4-iodobenzamide was prepared and evaluated for the potential imaging and staging of disseminated metastatic melanoma. Methods: [{sup 123}I]Iodonicotinamides were prepared by iododestannylation reactions using no-carrier-added iodine-123 and evaluated in vivo by biodistribution and competition studies and by single photon emission computed tomography (SPECT) imaging in black and albino nude mice bearing B16F0 murine melanotic and A375 human amelanotic melanoma tumours, respectively. Results: The iodonicotinamides displayed low-affinity binding for {sigma}{sub 1}-{sigma}{sub 2} receptors (K{sub i}>300 nM). In biodistribution studies in mice, N-(2-(diethylamino)ethyl)-5-[{sup 123}I]iodonicotinamide ([{sup 123}I]1) exhibited the fastest and highest uptake of the nicotinamide series in the B16F0 tumour at 1 h ({approx}8% ID/g), decreasing slowly over time. No uptake was observed in the A375 tumour. Clearance from the animals by urinary excretion was more rapid for N-alkyl-nicotinamides than for piperazinyl derivatives. At 1 h postinjection, the urinary excretion was 66% ID for [{sup 123}I]1, while the gastrointestinal tract amounted to 17% ID. Haloperidol was unable to reduce the uptake of [{sup 123}I]1 in pigmented mice, indicating that this uptake was likely due to an interaction with melanin. SPECT imaging of [{sup 123}I]1 in black mice bearing the B16F0 melanoma indicated that the radioactivity was predominately located in the tumour and eyes. No specific localisation was observed in nude mice bearing A375 amelanotic tumours. Conclusion: These findings suggest that [{sup 123}I]1, which displays high tumour uptake with rapid clearance from the body, could be a promising imaging agent for the detection of melanotic tumours.

  9. Cytotoxic evaluation of phenolic compounds from lichens against melanoma cells.

    Science.gov (United States)

    Brandão, Luiz Fabrício Gardini; Alcantara, Glaucia Braz; Matos, Maria de Fátima Cepa; Bogo, Danielle; Freitas, Deisy dos Santos; Oyama, Nathália Mitsuko; Honda, Neli Kika

    2013-01-01

    Atranorin, lichexanthone, and the (+)-usnic, diffractaic, divaricatic, perlatolic, psoromic, protocetraric, and norstictic acids isolated from the lichens Parmotrema dilatatum (VAIN.) HALE, Usnea subcavata MOTYKA, Usnea sp., Ramalina sp., Cladina confusa (SANT.) FOLMM. & AHTI, Dirinaria aspera HÄSÄNEN, and Parmotrema lichexanthonicum ELIASARO & ADLER were evaluated against UACC-62 and B16-F10 melanoma cells and 3T3 normal cells. Sulforhodamine B assay revealed significant cytotoxic activity in protocetraric, divaricatic, and perlatolic acids on UACC-62 cells (50% growth inhibitory concentration (GI(50)) 0.52, 2.7, and 3.3 µg/mL, respectively). Divaricatic and perlatolic acids proved the most active on B16-F10 cells (GI(50) 4.4, 18.0 µg/mL, respectively) and the most cytotoxic to 3T3 normal cells. Diffractaic, usnic, norstictic, and psoromic acids were cytotoxic to UACC-62 cells in the 24.7 to 36.6 µg/mL range, as were protocetraric and diffractaic acids to B16-F10 cells (GI(50) 24.0, 25.4 µg/mL, respectively). Protocetraric acid was highly selective (selectivity index (SI*) 93.3) against UACC-62 cells, followed by norstictic, perlatolic, psoromic, and divaricatic acids, while norstictic and divaricatic acids were more selective against B16-F10 cells. The high SI* value obtained for protocetraric acid on UACC-62 cells makes it a potential candidate for the study of melanomas in experimental models. Chemometric analysis was performed to evaluate the general behavior of the compounds against the cell lines tested. PMID:23207680

  10. Carbon ion induced DNA double-strand breaks in melanophore B16

    International Nuclear Information System (INIS)

    DNA double-strand breaks (DSBs) in melanophore B16 induced by plateau and extended Bragg peak of 75 MeV/u 12C6+ ions were studied by using a technique of inverse pulsed-field gel electrophoresis (PIGE). DNA fragment lengths were distributed in two ranges: the larger in 1.4 Mbp-3.2 Mbp and the smaller in less than 1.2 Mbp. It indicates that distribution of DNA fragments induced by heavy ion irradiation is not stochastic and there probably are sensitive sites to heavy ions in DNA molecules of B16. Percentage of DNA released from plug (PR) increased and trended towards a quasi-plateau ∝85% as dose increased. Content of the larger fragments decreased and flattened with increasing dose while content of the smaller ones increased and trended towards saturation. (orig.)

  11. Single-walled carbon nanotube-conjugated chemotherapy exhibits increased therapeutic index in melanoma

    Science.gov (United States)

    Chaudhuri, Padmaparna; Soni, Shivani; Sengupta, Shiladitya

    2010-01-01

    The incidence of malignant melanoma is increasing at an alarming rate globally. Poor prognosis and extraordinarily low survival rates of malignant melanoma necessitates the development of new chemotherapeutic strategies. An emerging approach is to harness nanotechnology to optimize the existing chemotherapies. In the present study we have demonstrated that the delivery of doxorubicin using a nanotechnology-based platform significantly reduces the systemic toxicity of the drug, keeping unchanged its therapeutic efficacy in a mouse melanoma tumor model. Specifically we modified single-walled carbon nanotubes (CNTs) to conjugate a doxorubicin prodrug via a carbamate linker that cleaves enzymatically to cause temporal release of the active drug. The CNT-doxorubicin conjugate (CNT-Dox) induced time-dependent cell death in B16-F10 melanoma cells in vitro. The nanoparticle was rapidly internalized into the lysosome of melanoma cells and was retained in the subcellular compartment for over 24 h. In an in vivo melanoma model, treatment with the nanotube-doxorubicin conjugate abrogated tumor growth without the systemic side-effects associated with free doxorubicin. Our studies demonstrate that a simple and versatile CNT-based nanovector can be harnessed for the delivery of chemotherapy to melanoma, with increased therapeutic index.

  12. Single-walled carbon nanotube-conjugated chemotherapy exhibits increased therapeutic index in melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Chaudhuri, Padmaparna; Soni, Shivani; Sengupta, Shiladitya, E-mail: shiladit@mit.edu [BWH-HST Center for Biomedical Engineering, Department of Medicine, Brigham and Women' s Hospital, Harvard Medical School, Harvard-MIT Division of Health Science and Technology, Cambridge, MA 02139 (United States)

    2010-01-15

    The incidence of malignant melanoma is increasing at an alarming rate globally. Poor prognosis and extraordinarily low survival rates of malignant melanoma necessitates the development of new chemotherapeutic strategies. An emerging approach is to harness nanotechnology to optimize the existing chemotherapies. In the present study we have demonstrated that the delivery of doxorubicin using a nanotechnology-based platform significantly reduces the systemic toxicity of the drug, keeping unchanged its therapeutic efficacy in a mouse melanoma tumor model. Specifically we modified single-walled carbon nanotubes (CNTs) to conjugate a doxorubicin prodrug via a carbamate linker that cleaves enzymatically to cause temporal release of the active drug. The CNT-doxorubicin conjugate (CNT-Dox) induced time-dependent cell death in B16-F10 melanoma cells in vitro. The nanoparticle was rapidly internalized into the lysosome of melanoma cells and was retained in the subcellular compartment for over 24 h. In an in vivo melanoma model, treatment with the nanotube-doxorubicin conjugate abrogated tumor growth without the systemic side-effects associated with free doxorubicin. Our studies demonstrate that a simple and versatile CNT-based nanovector can be harnessed for the delivery of chemotherapy to melanoma, with increased therapeutic index.

  13. Properties of kojic acid and curcumin: Assay on cell B16-F1

    Science.gov (United States)

    Sugiharto, Ariff, Arbakariya; Ahmad, Syahida; Hamid, Muhajir

    2016-03-01

    Ultra violet (UV) exposure and oxidative stress are casually linked to skin disorders. They can increase melanin synthesis, proliferation of melanocytes, and hyperpigmentation. It is possible that antioxidants or inhibitors may have a beneficial effect on skin health to reduce hyperpigmentation. In the last few years, a huge number of natural herbal extracts have been tested to reduce hyperpigmentation. The objective of this study was to determine and to compare of kojic acid and curcumin properties to viability cell B16-F1. In this study, our data showed that the viability of cell B16-F1 was 63.91% for kojic acid and 64.12% for curcumin at concentration 100 µg/ml. Further investigation assay of antioxidant activities, indicated that IC50 for kojic acid is 63.8 µg/ml and curcumin is 16.05 µg/ml. Based on the data, kojic acid and curcumin have potential antioxidant properties to reduce hyperpigmentation with low toxicity effect in cell B16-F1.

  14. Changes in intestinal microflora of Caenorhabditis elegans following Bacillus nematocida B16 infection.

    Science.gov (United States)

    Niu, Qiuhong; Zhang, Lin; Zhang, Keqin; Huang, Xiaowei; Hui, Fengli; Kan, Yunchao; Yao, Lunguang

    2016-01-01

    The effect of pathogenic bacteria on a host and its symbiotic microbiota is vital and widespread in the biotic world. The soil-dwelling opportunistic bacterium Bacillus nematocida B16 uses a "Trojan horse" mechanism to kill Caenorhabditis elegans. The alterations in the intestinal microflora that occur after B16 infection remain unknown. Here, we analyzed the intestinal bacteria presented in normal and infected worms. The gut microbial community experienced a complex change after B16 inoculation, as determined through marked differences in species diversity, structure, distribution and composition between uninfected and infected worms. Regardless of the worm's origin (i.e., from soil or rotten fruits), the diversity of the intestinal microbiome decreased after infection. Firmicutes increased sharply, whereas Proteobacteria, Actinobacteria, Cyanobacteria and Acidobacteria decreased to different degrees. Fusobacteria was only present 12 h post-infection. After 24 h of infection, 1228 and 1109 bacterial species were identified in the uninfected and infected groups, respectively. The shared species reached 21.97%. The infected group had a greater number of Bacillus species but a smaller number of Pediococcus, Halomonas, Escherichia and Shewanella species (P microbiota using C. elegans as the model species. PMID:26830015

  15. Recombinant snake venom cystatin inhibits the growth, invasion and metastasis of B16F10 cells and MHCC97H cells in vitro and in vivo.

    Science.gov (United States)

    Xie, Qun; Tang, Nanhong; Wan, Rong; Qi, Yuanlin; Lin, Xu; Lin, Jianyin

    2011-04-01

    Studies have shown that expression of snake venom cystatin (sv-cystatin) in mouse melanoma cells and human gastric carcinoma cells can inhibit their invasion and metastasis. To advance the research into the biological features and pharmaceutical applications of sv-cystatin, we investigated the expression of recombinant sv-cystatin in an optimized Pichia pastoris system. Approximately 5 mg/L of bioactive sv-cystatin was obtained with a purity of 95.08%. Kinetic analyses of recombinant sv-cystatin revealed highly effective inhibitory efficiency against papain (Ki = 2.67 nM). We further investigated the effects of recombinant sv-cystatin on the invasion and metastasis of B16F10 cells and MHCC97H cells in vitro and in vivo. Matrigel invasion assays showed significant inhibition of recombinant sv-cystatin on the tumor cells in vitro. For experimental lung colonization assays, C57BL/6 mice inoculated in the lateral tail vein with B16F10 cells were treated with three i.v. injections of recombinant sv-cystatin (25 and 50 mg/kg) 24 h before cell inoculation, and 2 h and 24 h after cell inoculation. Administration of recombinant sv-cystatin significantly suppressed the formation of lung tumor colonies. For spontaneous metastasis assays, MHCC97H cells were inoculated s.c. into nude mice. After 24 h, recombinant sv-cystatin was administered by i.p. injections at 25, 50 or 100 mg/kg once daily for 5 days. Administration of recombinant sv-cystatin significantly decreased the formation of lung tumor colonies. Taken together, recombinant sv-cystatin inhibits the invasion and metastasis of tumor cells in vitro and in vivo. These results may facilitate the future evaluation of the pharmaceutical applications of sv-cystatin.

  16. Amino acid alcohols: growth inhibition and induction of differentiated features in melanoma cells.

    Science.gov (United States)

    Landau, O; Wasserman, L; Deutsch, A A; Reiss, R; Panet, H; Novogrodsky, A; Nordenberg, J

    1993-05-14

    The effects of a series of D- and L-amino acid alcohols on the proliferation and phenotypic expression of B16 mouse melanoma cells were evaluated. B16 melanoma cells were incubated for different time intervals in the presence of D- or L-phenylalaninol (PHE), D- or L-alaninol (AL), D- or L-leucinol (LE), L-histidinol (HIS), L-tyrosinol (TYR) and L-methioninol (MET). All agents, including the D or L configuration, induced an anti-proliferative effect, although of considerably different magnitude. D-PHE was the most active growth inhibitor. The growth inhibitory effects were accompanied by phenotypic alterations, which included morphological changes and enhancement in the activities of NADPH cytochrome c reductase and tau-glutamyl transpeptidase. These phenotypic alterations correlated with the growth inhibitory effects of the different agents and seem to reflect a higher differentiated state. PMID:8099846

  17. 表达ESAT-6-gpi和IL-21的B16F10瘤苗的构建及其活性鉴定%Construction of B16F10 tumor vaccine expressing ESAT-6-gpi and IL-21 and identification of its biological activity

    Institute of Scientific and Technical Information of China (English)

    何向锋; 王净; 余方流; 赵枫姝; 张洪义; 曹文虎; 窦骏

    2011-01-01

    为构建膜表达糖基化磷脂酰肌醇(GPI)锚定的结核杆菌早期分泌靶抗原6 kD(ESAT-6 )和分泌IL-21的B16F10瘤苗并鉴定其活性,利用重叠PCR法构建pIRES-ESAT-6-gpi/IL-21重组质粒,以脂质体转染重组质粒到B16F10细胞,G418筛选出阳性克隆,用RT-PCR、免疫荧光、FCM和Western blot检测瘤苗细胞靶抗原表达,用瘤苗细胞培养上清刺激小鼠CD8+ T细胞,检测瘤苗所分泌IL-21的生物学活性.结果表明,pIRES-ESAT-6-gpi/IL-21重组质粒DNA测序正确,B16F10-ESAT-6-gpi/IL-21瘤苗细胞目的基因ESAT-6表达于瘤苗细胞表面,增殖能力未受外源基因导入影响,分泌的IL-21具有生物学活性,为研究膜表达ESAT-6和分泌表达IL-21瘤苗的抗瘤效应奠定了基础.%To construct B16F10 tumor vaccine that express the IL-21 and the 6 kDearly secreted antigenic target(ESAT-6) anchored by glycosylated phosphatidylinositol (GPI) on cell membrane and to identify its biological activity, plasmid recombination method based on overlap extension PCR was used to construct the eukaryotic expression vector for pIRES-ESAT-6-gpi/IL-21, then the melanoma B16F10 cells were transfected with the recombinant plasmid. The expressions of the fused protein and IL-21 were determined by RT-PCR. Immunofluorescence, FCM, Western blot assay, respectively. Mouse splenocytes were cultivated with supernatant from the tumor vaccine cells to detect the biological activity of IL-21 secreted by tumor vaccine cells. It was demonstrated that the ESAT-6 target molecule was expressed on the surface of B16F10-ESAT-6-gpi/IL-21 tumor vaccine cells, and the exogenous gene did not affect the proliferation of vaccine cells. Furthermore, the IL-21 secreted from vaccine cells has biological activity. The stable transfected B16F10-ESAT-6-gpi/IL-21 tumor vaccine cell line was successfully constructed and the vaccine could be used for further anti-tumor research.

  18. Bone marrow transplant - discharge

    Science.gov (United States)

    Transplant - bone marrow - discharge; Stem cell transplant - discharge; Hematopoietic stem cell transplant - discharge; Reduced intensity; Non-myeloablative transplant - discharge; Mini transplant - discharge; Allogenic bone marrow transplant - discharge; ...

  19. Macrophage-dependent clearance of systemically administered B16BL6-derived exosomes from the blood circulation in mice

    Directory of Open Access Journals (Sweden)

    Takafumi Imai

    2015-02-01

    Full Text Available Previous studies using B16BL6-derived exosomes labelled with gLuc–lactadherin (gLuc-LA, a fusion protein of Gaussia luciferase (a reporter protein and lactadherin (an exosome-tropic protein, showed that the exosomes quickly disappeared from the systemic circulation after intravenous injection in mice. In the present study, the mechanism of rapid clearance of intravenously injected B16BL6 exosomes was investigated. gLuc-LA-labelled exosomes were obtained from supernatant of B16BL6 cells after transfection with a plasmid DNA encoding gLuc-LA. Labelling was stable when the exosomes were incubated in serum. By using B16BL6 exosomes labelled with PKH26, a lipophilic fluorescent dye, it was demonstrated that PKH26-labelled B16BL6 exosomes were taken up by macrophages in the liver and spleen but not in the lung, while PKH26-labelled exosomes were taken up by the endothelial cells in the lung. Subsequently, gLuc-LA-labelled B16BL6 exosomes were injected into macrophage-depleted mice prepared by injection with clodronate-containing liposomes. The clearance of the intravenously injected B16BL6 exosomes from the blood circulation was much slower in macrophage-depleted mice than that in untreated mice. These results indicate that macrophages play important roles in the clearance of intravenously injected B16BL6 exosomes from the systemic circulation.

  20. Melanoma International Foundation

    Science.gov (United States)

    ... 501(c)(3) charity, also registered as a non-profit charity in the state of Pennsylvania, certificate #29498 © 2013 Melanoma International Foundation. All Rights Reserved. Privacy Policy | Terms of Use ...

  1. Drugs Approved for Melanoma

    Science.gov (United States)

    ... are not listed here. Drugs Approved for Melanoma Aldesleukin Cobimetinib Cotellic (Cobimetinib) Dabrafenib Dacarbazine DTIC-Dome (Dacarbazine) IL-2 (Aldesleukin) Imlygic (Talimogene Laherparepvec) Interleukin-2 (Aldesleukin) Intron A ( ...

  2. Proteomics in uveal melanoma.

    LENUS (Irish Health Repository)

    Ramasamy, Pathma

    2014-01-01

    Uveal melanoma is the most common primary intraocular malignancy in adults, with an incidence of 5-7 per million per year. It is associated with the development of metastasis in about 50% of cases, and 40% of patients with uveal melanoma die of metastatic disease despite successful treatment of the primary tumour. The survival rates at 5, 10 and 15 years are 65%, 50% and 45% respectively. Unlike progress made in many other areas of cancer, uveal melanoma is still poorly understood and survival rates have remained similar over the past 25 years. Recently, advances made in molecular genetics have improved our understanding of this disease and stratification of patients into low risk and high risk for developing metastasis. However, only a limited number of studies have been performed using proteomic methods. This review will give an overview of various proteomic technologies currently employed in life sciences research, and discuss proteomic studies of uveal melanoma.

  3. Genetics of Melanoma

    Directory of Open Access Journals (Sweden)

    Janet eWangari-Talbot

    2013-01-01

    Full Text Available Genomic variation is a trend observed in various human diseases including cancer. Genetic studies have set out to understand how and why these variations result in cancer, why some populations are predisposed to the disease, and also how genetics affect drug responses. The melanoma incidence has been increasing at an alarming rate worldwide. The burden posed by melanoma has made it a necessity to understand the fundamental signaling pathways involved in this deadly disease. Signaling cascades such as MAPK and PI3K/AKT have been shown to be crucial in the regulation of processes that are commonly dysregulated during cancer development such as aberrant proliferation, loss of cell cycle control, impaired apoptosis and altered drug metabolism. Understanding how these and other oncogenic pathways are regulated has been integral in our challenge to develop potent anti-melanoma drugs. With advances in technology and especially in next generation sequencing, we have been able to explore melanoma genomes and exomes leading to the identification of previously unknown genes with functions in melanomagenesis such as GRIN2A and PREX2. The therapeutic potential of these novel candidate genes is actively being pursued with some presenting as druggable targets while others serve as indicators of therapeutic responses. In addition, the analysis of the mutational signatures of melanoma tumors continues to cement the causative role of UV exposure in melanoma pathogenesis. It has become distinctly clear that melanomas from sun exposed skin areas have distinct mutational signatures including C to T transitions indicative of UV-induced damage. It is thus necessary to continue spreading awareness on how to decrease the risk factors of developing the disease while at the same time working for a cure. Given the large amount of information gained from these sequencing studies, it is likely that in the future, treatment of melanoma will follow a highly personalized route

  4. Primary Anorectal Melanoma: An Update

    Directory of Open Access Journals (Sweden)

    P Carcoforo, M.T Raiji, G.M Palini, M Pedriali, U Maestroni, G Soliani, A Detroia, M.V Zanzi, A.L Manna, J.G Crompton, R.C Langan, A Stojadinovic, I Avital

    2012-01-01

    Full Text Available The anorectum is a rare anatomic location for primary melanoma. Mucosal melanoma is a distinct biological and clinical entity from the more common cutaneous melanoma. It portrays worse prognosis than cutaneous melanoma, with distant metastases being the overwhelming cause of morbidity and mortality. Surgery is the treatment of choice, but significant controversy exists over the extent of surgical resection. We present an update on the state of the art of anorectal mucosal melanoma. To illustrate the multimodality approach to anorectal melanoma, we present a typical patient.

  5. Intravital Microscopy for Identifying Tumor Vessels in Patients With Stage IA-IV Melanoma That is Being Removed by Surgery

    Science.gov (United States)

    2016-01-13

    Recurrent Melanoma; Stage IA Skin Melanoma; Stage IB Skin Melanoma; Stage IIA Skin Melanoma; Stage IIB Skin Melanoma; Stage IIC Skin Melanoma; Stage IIIA Skin Melanoma; Stage IIIB Skin Melanoma; Stage IIIC Skin Melanoma; Stage IV Skin Melanoma

  6. Experimental stress analysis for four 24-in. ANSI standard B16.9 tees

    International Nuclear Information System (INIS)

    The experimental stress analysis and low cycle fatigue tests of four tees tested by Combustion Engineering, Inc. (E-E) under subcontract to Union Carbide Nuclear Division are described. These tests are part of the ORNL Design Criteria for Piping and Nozzles Program which is being conducted for the development of design criteria for nuclear power plant service piping components. The test assemblies were fabricated at C-E from commercially obtained ANSI B16.9 tees and matching diameter steel pipes welded to the tees, with suitable and closures and fixtures for applying the loads

  7. Treatment Options by Stage (Melanoma)

    Science.gov (United States)

    ... of Skin Cancer Skin Cancer Screening Research Melanoma Treatment (PDQ®)–Patient Version General Information About Melanoma Key ... Certain factors affect prognosis (chance of recovery) and treatment options. The prognosis (chance of recovery ) and treatment ...

  8. Fucoidan from Sargassum sp. and Fucus vesiculosus reduces cell viability of lung carcinoma and melanoma cells in vitro and activates natural killer cells in mice in vivo

    DEFF Research Database (Denmark)

    Ale, Marcel Tutor; Maruyama, Hiroko; Tamauchi, Hidekazu;

    2011-01-01

    Fucoidan is known to exhibit crucial biological activities, including anti-tumor activity. In this study, we examined the influence of crude fucoidan extracted from Sargassum sp. (MTA) and Fucus vesiculosus (SIG) on Lewis lung carcinoma cells (LCC) and melanoma B16 cells (MC). In vitro studies were...

  9. Dose estimation in B16 tumour bearing mice for future irradiation in the thermal column of the TRIGA reactor after B/Gd/LDL adduct infusion.

    Science.gov (United States)

    Protti, N; Ballarini, F; Bortolussi, S; Bruschi, P; Stella, S; Geninatti, S; Alberti, D; Aime, S; Altieri, S

    2011-12-01

    To test the efficacy of a new (10)B-vector compound, the B/Gd/LDL adduct synthesised at Torino University, in vivo irradiations of murine tumours are in progress at the TRIGA Mark II reactor of the Pavia University. A localised B16 melanoma tumour is generated in C57BL/6 mice and subsequently infused with the adduct. During the irradiation, the mouse will be put in a shield to protect the whole body except the tumour in the back-neck area. To optimise the treatment set-up, MCNP simulations were performed. A very simplified mouse model was built using MCNP geometry capabilities, as well as the geometry of the shield made of 99% (10)B enriched boric acid. A hole in the shield is foreseen in correspondence of the back-neck region. Many configurations of the shield were tested in terms of neutron flux, dose distribution and mean induced activity in the tumour region and in the radiosensitive organs of the mouse. In the final set-up, up to five mice can be treated simultaneously in the reactor thermal column and the neutron fluence in the tumour region for 10 min of irradiation is of about 5×10(12) cm(-2). PMID:21459587

  10. Mugil cephalus roe oil obtained by supercritical fluid extraction affects the lipid profile and viability in cancer HeLa and B16F10 cells.

    Science.gov (United States)

    Rosa, A; Piras, A; Nieddu, M; Putzu, D; Cesare Marincola, F; Falchi, A M

    2016-09-14

    We explored the changes in viability and lipid profile occurring in cancer cells, murine melanoma cells (B16F10 cells) and human cervical carcinoma cells (HeLa cells), when exposed to 24 h-treatments with an n-3 PUFA-rich oil obtained by supercritical extraction with CO2 from Mugil cephalus processed roe (bottarga). The composition of the major lipid classes of bottarga oil was determined by the (13)C NMR technique. Reversed-phase HPLC with DAD/ELSD detection was performed to analyze cells' total fatty acid profile and the levels of phospholipids, total/free cholesterol, triacylglycerols, and cholesteryl esters. Cell-based fluorescent measurements of intracellular membranes and lipid droplets were performed on bottarga oil-treated cells using the Nile red staining technique. The treatments of cancer cells with bottarga oil reduced the viability and affected the fatty acid profile, with a significant n-3 PUFA increase in treated cells. Mullet roe oil uptake modulated the cancer cell lipid composition, inducing a remarkable incorporation of health beneficial n-3 PUFA in the polar and neutral lipid fractions. Bottarga oil treatment influenced the synthesis of intracellular membranes and accumulation of cytoplasmic lipid droplets in cancer cells. PMID:27603212

  11. Um Melanoma “mascarado” Melanoma disfrazado A disguised Melanoma

    Directory of Open Access Journals (Sweden)

    Elias Ribeiro

    2012-08-01

    Full Text Available O melanoma é um tumor que se desenvolve como resultado da transformação maligna dos melanócitos, estimando-se a sua incidência global em 132.000 casos/ano. Este relato de caso reporta-se a um doente do sexo masculino com 70 anos, história de Diabetes Mellitus tipo 2 há dez anos e psoríase vulgar extensa há seis anos. Em aproximadamente um ano, este desenvolveu lesão ulcerada da região plantar do pé direito, que ao exame histológico revelou melanoma maligno, ulcerado, nível V de Clark, com 5,6 mm de espessura (Breslow. Foi submetido à exérese cirúrgica da lesão e biópsia de gânglio sentinela que foi negativa. O estadiamento inicial revelou evolução avançada do tumor primário (TNM IIC. Exames imagiológicos detetaram metastização gástrica, reclassificando a doença num estádio TNM IV. O melanoma maligno pode ser de difícil diagnóstico como se pode constatar neste caso em que uma ulceração plantar foi avaliada tardiamente, atrasando o diagnóstico de uma neoplasia grave e com elevada taxa de mortalidade. El melanoma es un tumor que se desarrolla como resultado de la transformación maligna de los melanocitos, estimándose su incidencia global en 132,000 casos/año. Este informe presenta a un paciente de sexo masculino de 70 años, con antecedentes de Diabetes Mellitus tipo 2 desde hace diez años y psoriasis vulgar extensa desde hace seis años. En aproximadamente un año el paciente desarrolló una lesión ulcerada en la región plantar del pie derecho, el examen histológico reveló un melanoma maligno, ulcerado, nivel V de Clark, de 5.6 mm de espesor (Breslow. Después de una escisión quirúrgica de la lesión, se realizó una biopsia de ganglio centinela que fue negativa. Las conclusiones iniciales revelaron una evolución avanzada del tumor primario (TNM IIC. Exámenes radiológicos detectaron una metástasis gástrica, reclasificando la enfermedad en una etapa TNM IV. El melanoma maligno puede ser de

  12. Antitumor effect of cytosine deaminase/5-fluorocytosine suicide gene therapy system mediated by Bifidobacterium infantis on melanoma

    Institute of Scientific and Technical Information of China (English)

    Cheng YI; Ying HUANG; Zhi-ying GUO; Shu-ren WANG

    2005-01-01

    Aim: To construct a Bifidobacterium infantis/CD targeting gene therapy system and observe the antitumor effect of cytosine deaminase/5-fluorocytosine (CD/5-FC) suicide gene therapy system mediated by Bifidobacterium infantis on melanoma in vitro and in vivo. Methods: A recombinant CD/pGEX- 1LamdaT plasmid was transfected into Bifidobacterium infantis by electroporation. Bifidobac terium infantis transfected by recombinant CD/pGEX-1LamdaT plasmid was in cubated with 5-FC anaerobically. Then the supernatant fluid was collected and added to melanoma B16-F10 cells to observe the killing effect for B16-F10 cells.Mice were inoculated with melanoma B 16-F10 cells to establish animal models.The mice were then injected with 5-FC and Bifidobacterium infantis transfected by recombinant CD/pGEX-1LamdaT plasmid. Results:Two segments of approxi mate 4.9 kb and 1.3 kb were extracted from the 6.2 kb recombinant plasmid, which were equal to the size of the pGEX-1LamdaT plasmid and CD gene, respectively.Sequencing results showed that the full length and sequence of nucleotide acid of the inserted gene in extracted recombinant plasmid was completely identical to the CD gene. In vitro, B 16-F10 cells treated by supernatant fluid were remarkably damaged morphologically, and the cell growth was significantly inhibited. Experi ments on the mice melanoma model showed that after treatment with a combination of transfected Bifidobacterium infantis and 5-FC, the tumor volume was significantly inhibited compared with controls. Conclusion: The foreign gene,CD gene, was correctly inserted into pGEX-1LambdaT plasmid and transferred into Bifidobacterium infantis. CD/5-FC suicide gene therapy system mediated by Bifidobacterium infantis demonstrated a good antitumor effect on melanoma in vitro and in vivo.

  13. Transplant services

    Science.gov (United States)

    ... may be done with a kidney- pancreas transplant . Liver transplant may be the only option for someone with liver disease that has led to liver failure. Lung transplant may replace one or both lungs. It may ...

  14. Liver transplant

    Science.gov (United States)

    ... transplant - series References Keefe EB. Hepatic failure and liver transplantation. In: Goldman L, Schafer AI, eds. Goldman's Cecil ... Elsevier; 2011:chap 157. Martin P, Rosen HR. Liver transplantation. In: Feldman M, Friedman LS, Brandt LJ, eds. ...

  15. Biofunctional Activities of Equisetum ramosissimum Extract: Protective Effects against Oxidation, Melanoma, and Melanogenesis

    Science.gov (United States)

    Li, Pin-Hui; Chiu, Yu-Pin; Shih, Chieh-Chih; Wen, Zhi-Hong; Ibeto, Laura Kaodichi; Huang, Shu-Hung; Chiu, Chien Chih; Ma, Dik-Lung; Leung, Chung-Hang; Chang, Yaw-Nan; Wang, Hui-Min David

    2016-01-01

    Equisetum ramosissimum, a genus of Equisetaceae, is a medicinal plant that can be separated into ethyl acetate (EA), dichloromethane (DM), n-hexane (Hex), methanol (MeOH), and water extracts. EA extract was known to have potent antioxidative properties, reducing power, DPPH scavenging activity, and metal ion chelating activity. This study compared these five extracts in terms of their inhibiting effects on three human malignant melanomas: A375, A375.S2, and A2058. MTT assay presented the notion that both EA and DM extracts inhibited melanoma growth but did not affect the viabilities of normal dermal keratinocytes (HaCaT) or fibroblasts. Western blot analyses showed that both EA and DM extracts induced overexpression of caspase proteins in all three melanomas. To determine their roles in melanogenesis, this study analyzed their in vitro suppressive effects on mushroom tyrosinase. All extracts except for water revealed moderate suppressive effects. None of the extracts affected B16-F10 cells proliferation. EA extract inhibited cellular melanin production whereas DM extract unexpectedly enhanced cellular pigmentation in B16-F10 cells. Data for modulations of microphthalmia-associated transcription factor, tyrosinase, tyrosinase-related protein 1, and tyrosinase-related protein 2 showed that EA extract inhibited protein expression mentioned above whereas DM extract had the opposite effect. Overall, the experiments indicated that the biofunctional activities of EA extract contained in food and cosmetics protect against oxidation, melanoma, and melanin production. PMID:27403230

  16. Melanoma targeting property of a Lu-177-labeled lactam bridge-cyclized alpha-MSH peptide

    OpenAIRE

    Guo, Haixun; Miao, Yubin

    2013-01-01

    The purpose of this study was to determine the melanoma targeting property of 177Lu-DOTA-GGNle-CycMSHhex in B16/F1 melanoma-bearing C57 mice. 177Lu-DOTA-GGNle-CycMSHhex exhibited high receptor-mediated melanoma uptake and fast urinary clearance. The tumor uptake of 177Lu-DOTA-GGNle-CycMSHhex was 20.25 ± 4.59 and 21.63 ± 6.27% ID/g at 0.5 and 2 h post-injection, respectively. Approximately 83% of injected dose cleared out the body via urinary system at 2 h post-injection. 177Lu-DOTA-GGNle-CycM...

  17. Melanoma Therapy with Rhenium-Cyclized Alpha Melanocyte Stimulating Hormone Peptide Analogs

    Energy Technology Data Exchange (ETDEWEB)

    Thomas P Quinn

    2005-11-22

    -DOTA-Re(Arg11)CCMSH and {sup 212}Pb-DOTA-Re(Arg11)CCMSH complexes were developed and synthesized to investigate its ability to target and deliver an effective dose to small melanoma tumors and metastatic deposits. Dosimetry calculations for {sup 188}Re-CCMSH and {sup 212}Pb/{sup 212}Bi[DOTA]-Re(Arg11)CCMSH were compared in the B16/F1 mouse melanoma flank tumor model to analyze the delivered dose to tumor and normal organs.

  18. Recombinant E.coli LLO/OVA Vaccination Effectively Inhibits Murine Melanoma Metastasis to Lung by CD8+T Cells Immunity

    Institute of Scientific and Technical Information of China (English)

    Man Xu; Ming-shen Dai; Can Mi

    2009-01-01

    Objective: To construct recombinant E.coli LLO/OVA and investigate its tumor metastatic inhibition effect in B16 OVA melanoma challenged mice.Methods: Recombinant E.coli LLO/OVA was constructed and the expression of listeriolysin O (LLO) and ovalbumin (OVA) of the vaccine was determined by coomassie brilliant blue staining and western blotting. After 3 subcutaneous injections of E.coli LLO/OVA, the percentages of CD3+CD4+T, CD4+CD25+T, CD3+CD8+T and OVA257(264 SIINFEKL specific CD8+T cells were determined by flow cytomytry, and the tumor metastatic inhibition effect in B16 OVA melanoma challenged mice was observed.Results: Recombinant E.coli LLO/OVA was successfully constructed, and the expression of LLO and OVA of the vaccine was confirmed. After 3 subcutaneous injections of E.coli LLO/OVA and E.coli OVA in mice, the percentages of CD3+CD4+T, CD4+CD25+T and CD3+CD8+T cells were equivalent in the two groups of mice. However, there were significantly more OVA257(264 SIINFEKL specific CD8+T cells in E.coli LLO/OVA vaccinated mice than that in E.coli OVA vaccinated mice. The prophylactic E.coli LLO/OVA vaccination effectively prevented the tumor metastasis to lungs in B16 OVA melanoma challenged mice. Depletion of CD8+T cells significantly impaired the tumor inhibition effect of the vaccine in B16 OVA challenged mice. The therapeutic vaccination of E.coli LLO/OVA significantly prevented melanoma metastasis to lungs in B16 OVA challenged mice too.Conclusion: Recombinant E.coli LLO/OVA vaccination is highly effective in inhibiting murine malignant melanoma metastasis by promoting CD8+T cell immunity.

  19. Intralesional injection of rose bengal induces a systemic tumor-specific immune response in murine models of melanoma and breast cancer.

    Directory of Open Access Journals (Sweden)

    Paul Toomey

    Full Text Available Intralesional (IL injection of PV-10 has shown to induce regression of both injected and non-injected lesions in patients with melanoma. To determine an underlying immune mechanism, the murine B16 melanoma model and the MT-901 breast cancer model were utilized. In BALB/c mice bearing MT-901 breast cancer, injection of PV-10 led to regression of injected and untreated contralateral subcutaneous lesions. In a murine model of melanoma, B16 cells were injected into C57BL/6 mice to establish one subcutaneous tumor and multiple lung lesions. Treatment of the subcutaneous lesion with a single injection of IL PV-10 led to regression of the injected lesion as well as the distant B16 melanoma lung metastases. Anti-tumor immune responses were measured in splenocytes collected from mice treated with IL PBS or PV-10. Splenocytes isolated from tumor bearing mice treated with IL PV-10 demonstrated enhanced tumor-specific IFN-gamma production compared to splenocytes from PBS-treated mice in both models. In addition, a significant increase in lysis of B16 cells by T cells isolated after PV-10 treatment was observed. Transfer of T cells isolated from tumor-bearing mice treated with IL PV-10 led to tumor regression in mice bearing B16 melanoma. These studies establish that IL PV-10 therapy induces tumor-specific T cell-mediated immunity in multiple histologic subtypes and support the concept of combining IL PV10 with immunotherapy for advanced malignancies.

  20. Gene transfer-applied BNCT (g-BNCT) for amelanotic melanoma in brain. Further upregulation of 10B uptake by cell modulation

    International Nuclear Information System (INIS)

    Our success in eradicating melanoma by single BNCT with BPA led to the next urgent theme, i.e. application of such BNCT for currently uncurable melanoma metastasis in brain. In order to establish 10B-BPA-BNCT for melanoma in brain, we have investigated the pharmacokinetics of BPA which is most critical factor for successful BNCT, in melanotic and amelanotic and further tyrosinase gene-transfected amelanotic melanoma proliferating in brain having blood-brain-barrier, as compared to melanoma proliferating in skin. We have established three implanted models for melanoma in brain: 1) A1059 cells, amelanotic melanoma, 2) B16B15b cells, melanotic melanoma cells, highly metastatic to brain, and 3) TA1059 cells, with active melanogenesis induced by tyrosinase gene transfection. We would like to report the results of comparative analysis of the BPA uptake ability in these melanoma cells in both brain and skin. Based on these findings, we are further investigating to enhance 10B-BPA uptake by not only g-BNCT but also by additional melanogenesis upregulating cell modulation. (author)

  1. Liposomes loaded with a STING pathway ligand, cyclic di-GMP, enhance cancer immunotherapy against metastatic melanoma.

    Science.gov (United States)

    Nakamura, Takashi; Miyabe, Hiroko; Hyodo, Mamoru; Sato, Yusuke; Hayakawa, Yoshihiro; Harashima, Hideyoshi

    2015-10-28

    Malignant melanomas escape immunosurveillance via the loss/down-regulation of MHC-I expression. Natural killer (NK) cells have the potential to function as essential effector cells for eliminating melanomas. Cyclic di-GMP (c-di-GMP), a ligand of the stimulator of interferon genes (STING) signal pathway, can be thought of as a new class of adjuvant against cancer. However, it is yet to be tested, because technologies for delivering c-di-GMP to the cytosol are required. Herein, we report that c-di-GMP efficiently activates NK cells and induces antitumor effects against malignant melanomas when loaded in YSK05 lipid containing liposomes, by assisting in the efficient delivery of c-di-GMP to the cytosol. The intravenous administration of c-di-GMP encapsulated within YSK05-liposomes (c-di-GMP/YSK05-Lip) into mice efficiently induced the production of type I interferon (IFN) as well as the activation of NK cells, resulting in a significant antitumor effect in a lung metastasis mouse model using B16-F10. This antitumor effect was dominated by NK cells. The infiltration of NK cells was observed in the lungs with B16-F10 melanomas. These findings indicate that the c-di-GMP/YSK05-Lip induces MHC-I non-restricted antitumor immunity mediated by NK cells. Consequently, c-di-GMP/YSK05-Lip represents a potentially new adjuvant system for use in immunotherapy against malignant melanomas.

  2. Metastatic melanoma imaging with an {sup 111}In-labeled lactam bridge-cyclized {alpha}-melanocyte-stimulating hormone peptide

    Energy Technology Data Exchange (ETDEWEB)

    Guo Haixun; Shenoy, Nalini; Gershman, Benjamin M.; Yang, Jianquan [College of Pharmacy, University of New Mexico, Albuquerque, NM 87131 (United States); Sklar, Larry A. [College of Pharmacy, University of New Mexico, Albuquerque, NM 87131 (United States); Cancer Research and Treatment Center, University of New Mexico, Albuquerque, NM 87131 (United States); Department of Pathology, University of New Mexico, Albuquerque, NM 87131 (United States); Miao Yubin [College of Pharmacy, University of New Mexico, Albuquerque, NM 87131 (United States); Cancer Research and Treatment Center, University of New Mexico, Albuquerque, NM 87131 (United States); Department of Dermatology, University of New Mexico, Albuquerque, NM 87131 (United States)], E-mail: ymiao@salud.unm.edu

    2009-04-15

    Introduction: The purpose of this study was to examine whether a novel lactam bridge-cyclized {sup 111}In-labeled 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-Gly-Glu-c[Lys-Nle-Glu-His-D-Phe-Arg-Trp-Gly-Arg-Pro-Val-Asp] {l_brace}DOTA-GlyGlu-CycMSH{r_brace} could be an effective imaging probe for metastatic melanoma detection. Methods: {sup 111}In-DOTA-GlyGlu-CycMSH was prepared and purified by reverse-phase high-performance liquid chromatography (RP-HPLC). The internalization and efflux of {sup 111}In-DOTA-GlyGlu-CycMSH were examined in B16/F10 melanoma cells. The biodistribution of {sup 111}In-DOTA-GlyGlu-CycMSH was determined in B16/F10 pulmonary metastatic melanoma-bearing and normal C57 mice. Pulmonary metastatic melanoma imaging was performed by small-animal single-photon emission computed tomography (SPECT)/CT (Nano-SPECT/CT) using {sup 111}In-DOTA-GlyGlu-CycMSH as an imaging probe and compared with 2-[{sup 18}F]fluoro-2-deoxy-D-glucose ([{sup 18}F]FDG) positron emission tomography (PET) imaging. Results: {sup 111}In-DOTA-GlyGlu-CycMSH was readily prepared with greater than 95% radiolabeling yield. {sup 111}In-DOTA-GlyGlu-CycMSH displayed rapid internalization and extended efflux in B16/F10 cells. {sup 111}In-DOTA-GlyGlu-CycMSH exhibited significantly (P<.05) higher uptakes (2.00{+-}0.74%ID/g at 2 h post-injection and 1.83{+-}0.12%ID/g at 4 h post-injection) in metastatic melanoma-bearing lung than that in normal lung (0.08{+-}0.08%ID/g and 0.05{+-}0.05%ID/g at 2 and 4 h post-injection, respectively). The activity accumulation in normal organs was low (<0.5%ID/g) except for the kidneys 2 and 4 h post-injection. B16/F10 pulmonary melanoma metastases were clearly visualized with {sup 111}In-DOTA-GlyGlu-CycMSH 2 h post-injection rather than with [{sup 18}F]FDG 1 h post-injection. Conclusions: {sup 111}In-DOTA-GlyGlu-CycMSH exhibited favorable metastatic melanoma-targeting and -imaging properties, highlighting its potential as an effective imaging

  3. Pancreas Transplantation

    Science.gov (United States)

    ... by Mail Close www.diabetes.org > Living With Diabetes > Treatment and Care > Transplantation Share: Print Page Text Size: A A ... cc-kidneys-nephropathy, In this section Living With Diabetes Treatment and Care Transplantation Islet Transplantation Pancreas Transplantation Kidney Replacement Therapy ...

  4. Effective melanoma immunotherapy in mice by the skin-depigmenting agent monobenzone and the adjuvants imiquimod and CpG.

    Directory of Open Access Journals (Sweden)

    Jasper G van den Boorn

    Full Text Available BACKGROUND: Presently melanoma still lacks adequate treatment options for metastatic disease. While melanoma is exceptionally challenging to standard regimens, it is suited for treatment with immunotherapy based on its immunogenicity. Since treatment-related skin depigmentation is considered a favourable prognostic sign during melanoma intervention, we here aimed at the reverse approach of directly inducing vitiligo as a shortcut to effective anti-melanoma immunity. METHODOLOGY AND PRINCIPAL FINDINGS: We developed an effective and simple to use form of immunotherapy by combining the topical skin-bleaching agent monobenzone with immune-stimulatory imiquimod cream and cytosine-guanine oligodeoxynucleotides (CpG injections (MIC therapy. This powerful new approach promptly induced a melanoma antigen-specific immune response, which abolished subcutaneous B16.F10 melanoma growth in up to 85% of C57BL/6 mice. Importantly, this regimen induced over 100 days of tumor-free survival in up to 60% of the mice, and forcefully suppressed tumor growth upon re-challenge either 65- or 165 days after MIC treatment cessation. CONCLUSIONS: MIC therapy is effective in eradicating melanoma, by vigilantly incorporating NK-, B- and T cells in its therapeutic effect. Based on these results, the MIC regimen presents a high-yield, low-cost and simple therapy, readily applicable in the clinic.

  5. Stimulation of lymphocyte anti-melanoma activity by co-cultured macrophages activated by complex homeopathic medication

    International Nuclear Information System (INIS)

    Melanoma is the most aggressive form of skin cancer, and the most rapidly expanding cancer in terms of worldwide incidence. Chemotherapeutic approaches to treat melanoma have been uniformly disappointing. A Brazilian complex homeopathic medication (CHM), used as an immune modulator, has been recommended for patients with depressed immune systems. Previous studies in mice have demonstrated that the CHM activates macrophages, induces an increase in the number of leukocytes and improves the murine response against Sarcoma-180. Here we studied the interaction of mouse lymph node lymphocytes, co-cultured in vitro with macrophages in the presence or absence of the CHM, with B16F10 melanoma cells. Lymphocytes co-cultured with macrophages in the presence of the CHM had greater anti-melanoma activity, reducing melanoma cell density and increasing the number of lysed tumor cells. There was also a higher proportion of activated (CD25+) lymphocytes with increased viability. Overall, lymphocytes activated by treatment destroyed growing cancer cells more effectively than control lymphocytes. Co-culture of macrophages with lymphocytes in the presence of the CHM enhanced the anti-cancer performance of lymphocytes against a very aggressive lineage of melanoma cells. These results suggest that non-toxic therapies using CHMs are a promising alternative approach to the treatment of melanomas. In addition, they are attractive combination-therapy candidates, which may enhance the efficacy of conventional medicines by improving the immune response against tumor cells

  6. Pancreatic transplantation

    Directory of Open Access Journals (Sweden)

    Åke Andrén-Sandberg

    2010-01-01

    Full Text Available A pancreas transplant is a surgical procedure to place a healthy pancreas from a donor into a patient whose pancreas no longer functions properly. Exocrine pancreas transplantation remains the standard treatment of choice for patients with diabetes mellitus complicated by end-stage renal disease. The use of pancreas transplantation for type 2 diabetes mellitus is an emerging concept. A pancreas transplant is often done in conjunction with a kidney transplant. Even if pancreas transplantation provides the best glycemic control option for diabetes mellitus, it is associated with significant morbidities related to infectious disease. The present article provides with a review of pancreatic transplantation.

  7. Malignant melanoma of nose

    OpenAIRE

    Kundu, I. N.; Haldar, B.; Saha, A. K.

    2001-01-01

    Malignant melanoma (MM) is one of the uncommon malignancies of the nose. We present an unusually big proliferative like MM in the vestibule of the nose. Malignancy of nose constitutes less than 1% of all malignancies (3% of head & neck tumour). MM however contributes only 2% of all malignant neoplasms of the nose (Moore & Martin. 1955).

  8. Chemotherapy for Melanoma.

    Science.gov (United States)

    Wilson, Melissa A; Schuchter, Lynn M

    2016-01-01

    Prior to the recent therapeutic advances, chemotherapy was the mainstay of treatment options for advanced-stage melanoma. A number of studies have investigated various chemotherapy combinations in order to expand on the clinical responses achieved with single-agent dacarbazine, but these have not demonstrated an improvement in overall survival. Similar objective responses were observed with the combination of carboplatin and paclitaxel as were seen with single-agent dacarbazine. The combination of chemotherapy and immunotherapy, known as biochemo-therapy, has shown high clinical responses; however, biochemo-therapy has not been shown to improve overall survival and resulted in increased toxicities. In contrast, palliation and long-term responses have been observed with localized treatment with isolated limb perfusion or infusion in limb-isolated disease. Although new, improved therapeutic options exist for first-line management of advanced-stage melanoma, chemotherapy may still be important in the palliative treatment of refractory, progressive, and relapsed melanoma. We review the various chemotherapy options available for use in the treatment and palliation of advanced-stage melanoma, discuss the important clinical trials supporting the treatment recommendations, and focus on the clinical circumstances in which treatment with chemotherapy is useful.

  9. Melanoma Risk Prediction Models

    Science.gov (United States)

    Developing statistical models that estimate the probability of developing melanoma cancer over a defined period of time will help clinicians identify individuals at higher risk of specific cancers, allowing for earlier or more frequent screening and counseling of behavioral changes to decrease risk.

  10. Spice Blocks Melanoma Growth

    Science.gov (United States)

    Science Teacher, 2005

    2005-01-01

    Curcumin, the pungent yellow spice found in both turmeric and curry powders, blocks a key biological pathway needed for development of melanoma and other cancers, according to a study that appears in the journal Cancer. Researchers from The University of Texas M. D. Anderson Cancer Center demonstrate how curcumin stops laboratory strains of…

  11. Melanoma risk prediction models

    Directory of Open Access Journals (Sweden)

    Nikolić Jelena

    2014-01-01

    Full Text Available Background/Aim. The lack of effective therapy for advanced stages of melanoma emphasizes the importance of preventive measures and screenings of population at risk. Identifying individuals at high risk should allow targeted screenings and follow-up involving those who would benefit most. The aim of this study was to identify most significant factors for melanoma prediction in our population and to create prognostic models for identification and differentiation of individuals at risk. Methods. This case-control study included 697 participants (341 patients and 356 controls that underwent extensive interview and skin examination in order to check risk factors for melanoma. Pairwise univariate statistical comparison was used for the coarse selection of the most significant risk factors. These factors were fed into logistic regression (LR and alternating decision trees (ADT prognostic models that were assessed for their usefulness in identification of patients at risk to develop melanoma. Validation of the LR model was done by Hosmer and Lemeshow test, whereas the ADT was validated by 10-fold cross-validation. The achieved sensitivity, specificity, accuracy and AUC for both models were calculated. The melanoma risk score (MRS based on the outcome of the LR model was presented. Results. The LR model showed that the following risk factors were associated with melanoma: sunbeds (OR = 4.018; 95% CI 1.724- 9.366 for those that sometimes used sunbeds, solar damage of the skin (OR = 8.274; 95% CI 2.661-25.730 for those with severe solar damage, hair color (OR = 3.222; 95% CI 1.984-5.231 for light brown/blond hair, the number of common naevi (over 100 naevi had OR = 3.57; 95% CI 1.427-8.931, the number of dysplastic naevi (from 1 to 10 dysplastic naevi OR was 2.672; 95% CI 1.572-4.540; for more than 10 naevi OR was 6.487; 95%; CI 1.993-21.119, Fitzpatricks phototype and the presence of congenital naevi. Red hair, phototype I and large congenital naevi were

  12. Timosaponin AIII inhibits melanoma cell migration by suppressing COX-2 and in vivo tumor metastasis.

    Science.gov (United States)

    Kim, Ki Mo; Im, A-Rang; Kim, Seung Hyung; Hyun, Jin Won; Chae, Sungwook

    2016-02-01

    Melanoma is the leading cause of death from skin disease, due in large part to its propensity to metastasize. We examined the effects of timosaponin AIII, a compound isolated from Anemarrhena asphodeloides Bunge, on melanoma cancer cell migration and the molecular mechanisms underlying these effects using B16-F10 and WM-115 melanoma cells lines. Overexpression of COX-2, its metabolite prostaglandin E2 (PGE2), and PGE2 receptors (EP2 and EP4) promoted cell migration in vitro. Exposure to timosaponin AIII resulted in concentration-dependent inhibition of cell migration, which was associated with reduced levels of COX-2, PGE2, and PGE2 receptors. Transient transfection of COX-2 siRNA also inhibited cell migration. Exposure to 12-O-tetradecanoylphorbal-13-acetate enhanced cell migration, whereas timosaponin AIII inhibited 12-O-tetradecanoylphorbal-13-acetate-induced cell migration and reduced basal levels of EP2 and EP4. Moreover, timosaponin AIII inhibited activation of nuclear factor-kappa B (NF-κB), an upstream regulator of COX-2 in B16-F10 cells. Consistent with our in vitro findings, in vivo studies showed that timosaponin AIII treatment significantly reduced the total number of metastatic nodules in the mouse lung and improved histological alterations in B16-F10-injected C57BL/6 mice. In addition, C57BL/6 mice treated with timosaponin AIII showed reduced expression of COX-2 and NF-κB in the lung. Together, these results indicate that timosaponin AIII has the capacity to inhibit melanoma cell migration, an essential step in the process of metastasis, by inhibiting expression of COX-2, NF-κB, PGE2, and PGE2 receptors.

  13. Melanoma maligno conjuntival

    Directory of Open Access Journals (Sweden)

    Gustavo Amorim Novais

    2012-08-01

    Full Text Available INTRODUÇÃO: Os tumores melanocíticos conjuntivais compreendem lesões que podem variar desde lesões benignas como os nevos conjuntivais, lesões pré-cancerosas como melanose adquirida primária com atipia até o melanoma maligno conjuntival. O reconhecimento das características clinicas destas lesões e seu diagnóstico preciso permitem o tratamento adequado, contribuindo para a redução da morbidade e mortalidade associados ao melanoma de conjuntiva. OBJETIVO: Revisão das características clinicas, diagnóstico e modalidades de tratamento clínico e cirúrgico das lesões precursoras (Nevos e melanose adquirida primária e do maligno conjuntival. MÉTODOS: Revisão de literatura através de pesquisa no banco de dados MEDLINE, PUBMED, LILACS e SciELO no período de 1980 a 2011. As palavras-chave utilizadas, individualmente ou em conjunto, foram: "conjunctival melanoma", "primary acquired melanosis", "nevi", "treatment", "chemotherapy", "recurrence", "metastasis" e "mortality". RESULTADO: Características clínicas que permitem o diagnóstico do melanoma conjuntival e sua diferenciação de outras lesões pigmentadas conjuntivais foram consideradas. O estadiamento clínico e patológico, assim como modalidades de tratamento para o melanoma maligno foram revisadas. CONCLUSÕES: Pacientes portadores de lesões pigmentadas conjuntivais devem ser avaliados por um oftalmologista especialista. A história oftalmológica, a história familiar de melanoma, e características clínicas da lesão necessitam de cuidadosa avaliação, incluindo-se a determinação do risco de malignidade. A documentação fotográfica deve ser realizada. O tratamento clínico e planejamento cirúrgico devem ser baseados na suspeita clinica. A análise histopatológica por patologista experiente é fundamental para orientação do tratamento e para identificação de fatores prognósticos, principalmente em casos de doença maligna.

  14. Influence of rosmarinic acid and Salvia officinalis extracts on melanogenesis of B16F10 cells

    Directory of Open Access Journals (Sweden)

    Karina B. Oliveira

    2012-01-01

    Full Text Available Melanin is a photoprotective skin pigment, and pathologies characterized by hypo or hyperpigmentation are common. New compounds that regulate melanogenesis are, therefore, opportune, and many natural products with this property, as polyphenols, have been described. Salvia officinalis L., Lamiaceae, is a widely used food spice that contains high amounts of phenol derivates, including rosmarinic acid. The aim of this work was to evaluate the contribution of rosmarinic acid in the melanogenic activity of sage extracts. Fluid and aqueous extracts of sage and purified rosmarinic acid were assayed for B16F10 cytotoxicity and, then, evaluated on melanin production and tyrosinase activity. While sage extracts showed a concentration-dependent ability to significantly increase melanin production without necessarily changing the enzymatic activity, rosmarinic acid showed a dual behavior on melanogenesis, increasing melanin biosynthesis and tyrosinase activity at low concentrations and decreasing it at higher levels. Rosmarinic acid may collaborate with sage extracts activity on melanogenesis, although other compounds may be involved. This is the first time that a dual action of rosmarinic acid on melanogenesis is reported, which may be useful in further studies for therapeutic formulations to treat skin pigmentation disorders.

  15. Influence of rosmarinic acid and Salvia officinalis extracts on melanogenesis of B16F10 cells

    Directory of Open Access Journals (Sweden)

    Karina B. Oliveira

    2013-04-01

    Full Text Available Melanin is a photoprotective skin pigment, and pathologies characterized by hypo or hyperpigmentation are common. New compounds that regulate melanogenesis are, therefore, opportune, and many natural products with this property, as polyphenols, have been described. Salvia officinalis L., Lamiaceae, is a widely used food spice that contains high amounts of phenol derivates, including rosmarinic acid. The aim of this work was to evaluate the contribution of rosmarinic acid in the melanogenic activity of sage extracts. Fluid and aqueous extracts of sage and purified rosmarinic acid were assayed for B16F10 cytotoxicity and, then, evaluated on melanin production and tyrosinase activity. While sage extracts showed a concentration-dependent ability to significantly increase melanin production without necessarily changing the enzymatic activity, rosmarinic acid showed a dual behavior on melanogenesis, increasing melanin biosynthesis and tyrosinase activity at low concentrations and decreasing it at higher levels. Rosmarinic acid may collaborate with sage extracts activity on melanogenesis, although other compounds may be involved. This is the first time that a dual action of rosmarinic acid on melanogenesis is reported, which may be useful in further studies for therapeutic formulations to treat skin pigmentation disorders.

  16. Forging of Naval Brass (ASTM B16) - Finite Element Analysis using Ls Dyna

    Science.gov (United States)

    Subha Sankari, T.; Sangavi, S.; Paneerselvam, T.; Venkatraman, R.; Venkatesan, M.

    2016-09-01

    Forging is one of the important manufacturing process in which products like connecting rod, transmission shaft, clutch hubs and gears are produced. Finite element analysis (FEA) in forming techniques is of recent interest for the optimal design and determination of right manufacturing forming process. The data from the numerical results can help in providing the information for selecting the ideal process conditions. Thus aside from experimental values, simulation by the finite element analysis software's such as LS DYNA can be used for the analysis of strain distribution in forging processes. In the present work, Finite element simulation of open die forging of naval brass (ASTM B16) is done at an optimal temperature. An advanced multi physics simulation software package by the Livermore software technology cooperation LSTC - LS DYNA is utilized for the simulation of forging process. For the forging validation, experiment is conducted with a cylindrical billet having height 45 mm and diameter of 40mm. The numerical results are compared with that of experimental results carried out at the same temperature and dimensions for validation. The distribution of strain is analyzed. Energy analysis due to impact load is detailed. The simulation results are found to be in good agreement with the experimental results.

  17. Multidrug resistance in ocular melanoma.

    OpenAIRE

    McNamara, M.; Clynes, M.; Dunne, B; NicAmhlaoibh, R; Lee, W. R.; Barnes, C; Kennedy, S M

    1996-01-01

    AIMS/BACKGROUND: Metastatic disease in patients with ocular melanoma is resistant to chemotherapy. One of the main mechanisms of modulating multidrug resistance is the expression of the multidrug resistance gene 1 (MDR1) product (p-glycoprotein) by tumour cells. The purpose of this study was to evaluate the frequency of expression of the MDR1 gene in ocular melanoma whose primary treatment was surgical excision or enucleation. METHODS: Twelve recent ocular melanomas were received fresh, snap ...

  18. Hair transplantation.

    Science.gov (United States)

    Avram, Marc R

    2012-12-01

    Hair transplantation is a purely dermatologic surgical procedure that dermatologists should be able to perform in appropriate candidates with hair loss. Hair transplantation techniques performed in the 1960s through the 1990s utilized large grafts that created an unfortunate public image of unnatural-appearing transplanted hair. Over the last 15 years, hair transplantation has been performed using follicular units to create consistently natural-looking transplanted hair in both men and women. This article provides an overview of candidate selection and state-of-the-art techniques for performing hair transplantation. PMID:23409484

  19. Host kinin B1 receptor plays a protective role against melanoma progression.

    Science.gov (United States)

    Maria, Andrea G; Dillenburg-Pilla, Patrícia; Reis, Rosana I; Floriano, Elaine M; Tefé-Silva, Cristiane; Ramos, Simone G; Pesquero, João B; Nahmias, Clara; Costa-Neto, Claudio M

    2016-01-01

    Melanoma is a very aggressive tumor that arises from melanocytes. Late stage and widely spread diseases do not respond to standard therapeutic approaches. The kallikrein-kinin system (KKS) participates in biological processes such as vasodilatation, pain and inflammatory response. However, the role of KKS in tumor formation and progression is not completely understood. The role of the host kinin B1 receptor in melanoma development was evaluated using a syngeneic melanoma model. Primary tumors and metastasis were respectively induced by injecting B16F10 melanoma cells, which are derived from C57BL/6 mice, subcutaneously or in the tail vein in wild type C57BL/6 and B1 receptor knockout mice (B1(-/-)). Tumors developed in B1(-/-) mice presented unfavorable prognostic factors such as increased incidence of ulceration, higher levels of IL-10, higher activation of proliferative pathways such as ERK1/2 and Akt, and increased mitotic index. Furthermore, in the metastasis model, B1(-/-) mice developed larger metastatic colonies in the lung and lower CD8(+)immune effector cells when compared with WT animals. Altogether, our results provide evidences that B1(-/-) animals developed primary tumors with multiple features associated with poor prognosis and unfavorable metastatic onset, indicating that the B1 receptor may contribute to improve the host response against melanoma progression. PMID:26898917

  20. Characterization of the melanoma brain metastatic niche in mice and humans

    International Nuclear Information System (INIS)

    Brain metastases occur in 15% of patients with melanoma and are associated with a dismal prognosis. Here, we investigate the architectural phenotype and stromal reaction of melanoma brain metastasis in mice and humans. A syngeneic, green fluorescence protein (GFP)-expressing murine B16-F1 melanoma clone was introduced via intracardiac injection, and was examined in vivo in comparison with human specimens. Immunofluorescence analyses of the brain metastases revealed that F4/80+ macrophages/microglia were most abundant at the tumor front, but rare in its core, where they were found only around blood vessels (P = 0.01). Similar pattern of infiltration was found in CD3+ T cells (P < 0.01). Infiltrating T cells were prominently CD4+ compared with CD8+ T cells (P < 0.001). Blood vessels (CD31+) were less abundant at the tumor front than in its center (12 ± 1 vs. 4 ± 0.6 vessels per high-power field [HPF], P < 0.001). In contrast, there were few vessels at the tumor front, but their diameter was significantly larger at the front (8236 μm2 vs. 4617 μm2 average cross-sectional area, P < 0.005). This is the first comparative analysis of melanoma brain metastases showing similar stromal reaction in murine models and human specimens. Our results validate the utility of this murine model of melanoma brain metastases for investigating the mechanism of the human disease

  1. Role of Ets-1 in fibronectin-derived heparin-binding domain polypeptides alleviating melanoma cell invasiveness and chemoresistance.

    Science.gov (United States)

    Tang, Nanhong; Wang, Xiaoqian; Huang, Tao; Wu, Yong; Chen, Yuanzhong

    2014-07-01

    In this study, we observed that rhFNHN29 and rhFNHC36, two recombinant heparin-binding domain polypeptides of fibronectin, suppressed adhesion and invasion of B16F10 and A375 melanoma cells mediated by integrin αv and α2 in a dose-dependent manner. Combined with low-concentration epirubicin (EPI), rhFNHN29 or rhFNHC36 exhibited a synergistic inhibition on the viability and metastasis of B16F10 cells. Moreover, in the presence of high-concentration rhFNHN29 or rhFNHC36, the Ets-1 activity and the expression of p-FAK, p-Erk1/2 and Ets-1 were notably downregulated in B16F10 cells. Ets-1 is one of the central regulatory links for rhFNHN29 and rhFNHC36 to suppress the adhesion and invasion of melanoma cells. Combining rhFNHN29 or rhFNHC36 with EPI may be a good way to alleviate invasiveness or chemoresistance in melanoma.

  2. Inhibition of B16BL6 tumor progression by coadministration of recombinant angiostatin K1-3 and endostatin genes with cationic liposomes.

    Science.gov (United States)

    Kim, Keun Sik; Kim, Hong Sung; Park, Jin Seu; Kwon, Young Guen; Park, Yong Serk

    2004-06-01

    Transfection of the antiangiogenic angiostatin and endostatin genes was shown to be an alternative to high-dose administration of angiostatin or endostatin proteins for cancer therapy. We have systematically investigated whether coadministration of the mouse angiostatin kringle 1-3 gene (pFLAG-AngioK1/3) and the endostatin gene (pFLAG-Endo) complexed with cationic liposomes exhibits enhanced therapeutic efficacy. In vitro, the coexpressed mixture of angiostatin K1-3 and endostatin more effectively reduced angiogenesis in chorioallantoic membranes than either angiostatin K1-3 or endostatin alone. In vivo, subcutaneous co-administration of pFLAG-AngioK1/3 and pFLAG-Endo lipoplexes more effectively inhibited vascularization in Matrigel plugs implanted in mice than either one alone. Additionally, subcutaneous administration of these genes inhibited the growth and formation of pulmonary metastases of B16BL6 melanoma cells in mice. Compared to treatment with an empty vector, treatment with pFLAG-AngioK1/3 plus pFLAG-Endo inhibited 81% of tumor growth, while treatment with pFLAG-AngioK1/3 or pFLAG-Endo inhibited tumor growth 70 and 69%, respectively. Cotreatment with the two plasmids after primary tumor excision induced a 90% inhibition of pulmonary metastases versus 79% for pFLAG-AngioK1/3 or 80% for pFLAG-Endo individually. These results suggest that combined administration of angiostatin K1-3 and endostatin genes complexed with cationic liposomes may be an innovated antiangiogenic strategy for cancer therapy. PMID:15118757

  3. Dehydroleucodine inhibits tumor growth in a preclinical melanoma model by inducing cell cycle arrest, senescence and apoptosis.

    Science.gov (United States)

    Costantino, Valeria V; Lobos-Gonzalez, Lorena; Ibañez, Jorge; Fernandez, Dario; Cuello-Carrión, F Darío; Valenzuela, Manuel A; Barbieri, Manuel A; Semino, Silvana N; Jahn, Graciela A; Quest, Andrew F G; Lopez, Luis A

    2016-03-01

    Malignant melanoma represents the fastest growing public health risk of all cancer types worldwide. Several strategies and anti-cancer drugs have been used in an effort to improve treatments, but the development of resistance to anti-neoplastic drugs remains the major cause of chemotherapy failure in melanomas. Previously, we showed that the sesquiterpene lactone, dehydroleucodine (DhL), promotes the accumulation of DNA damage markers, such as H2AX and 53BP1, in human tumor cells. Also DhL was shown to trigger either cell senescence or apoptosis in a concentration-dependent manner in HeLa and MCF7 cells. Here, we evaluated the effects of DhL on B16F0 mouse melanoma cells in vitro and in a pre-clinical melanoma model. DhL inhibited the proliferation of B16F0 cells by inducing senescence or apoptosis in a concentration-dependent manner. Also, DhL reduced the expression of the cell cycle proteins cyclin D1 and B1 and the inhibitor of apoptosis protein, survivin. In melanomas generated by subcutaneous injection of B16F0 cells into C57/BL6 mice, the treatment with 20 mg DhL /Kg/day in preventive, simultaneous and therapeutic protocols reduced tumor volumes by 70%, 60% and 50%, respectively. DhL treatments reduced the number of proliferating, while increasing the number of senescent and apoptotic tumor cells. To estimate the long-term effects of DhL, a mathematical model was applied to fit experimental data. Extrapolation beyond experimental time points revealed that DhL administration following preventive and therapeutic protocols is predicted to be more effective than simultaneous treatments with DhL in restricting tumor growth. PMID:26718258

  4. Local immunosuppressive microenvironment enhances migration of melanoma cells to lungs in DJ-1 knockout mice.

    Directory of Open Access Journals (Sweden)

    Chia-Hung Chien

    Full Text Available DJ-1 is an oncoprotein that promotes survival of cancer cells through anti-apoptosis. However, DJ-1 also plays a role in regulating IL-1β expression, and whether inflammatory microenvironment built by dysregulated DJ-1 affects cancer progression is still unclear. This study thus aimed to compare the metastatic abilities of melanoma cells in wild-type (WT and DJ-1 knockout (KO mice, and to check whether inflammatory microenvironment built in DJ-1 KO mice plays a role in migration of cancer cells to lungs. First, B16F10 melanoma cells (at 6 × 10(4 were injected into the femoral vein of mice, and formation of lung nodules, levels of lung IL-1β and serum cytokines, and accumulation of myeloid-derived suppressor cells (MDSCs were compared between WT and DJ-1 KO mice. Second, the cancer-bearing mice were treated with an interleukin-1 beta (IL-1β neutralizing antibody to see whether IL-1β is involved in the cancer migration. Finally, cultured RAW 264.7 macrophage and B16F10 melanoma cells were respectively treated with DJ-1 shRNA and recombinant IL-1β to explore underlying molecular mechanisms. Our results showed that IL-1β enhanced survival and colony formation of cultured melanoma cells, and that IL-1β levels were elevated both in DJ-1 KO mice and in cultured macrophage cells with DJ-1 knockdown. The elevated IL-1β correlated with higher accumulation of immunosuppressive MDSCs and formation of melanoma module in the lung of DJ-1 KO mice, and both can be decreased by treating mice with IL-1β neutralizing antibodies. Taken together, these results indicate that immunosuppressive tissue microenvironment built in DJ-1 KO mice can enhance lung migration of cancer, and IL-1β plays an important role in promoting the cancer migration.

  5. Topical and Targeted Delivery of siRNAs to Melanoma Cells Using a Fusion Peptide Carrier.

    Science.gov (United States)

    Ruan, Renquan; Chen, Ming; Sun, Sijie; Wei, Pengfei; Zou, Lili; Liu, Jing; Gao, Dayong; Wen, Longping; Ding, Weiping

    2016-01-01

    Topical application of siRNAs through the skin is a potentially effective strategy for the treatment of melanoma tumors. In this study, we designed a new and safe fusion peptide carrier SPACE-EGF to improve the skin and cell penetration function of the siRNAs and their targeting ability to B16 cells, such that the apoptosis of B16 cells can be induced. The results show that the carrier is stable and less toxic. The EGF motif does not affect the skin and cell penetration function of the SPACE. Because EGF can strongly bind EGFR, which is overexpressed in cancer cells, the targeting ability of the SPACE-EGF-siRNA complex is increased. In vitro experiments indicate that GAPDH siRNAs conjugated with SPACE-EGF can significantly reduce the GAPDH concentration in B16 cells, and c-Myc siRNAs can cause the gene silencing of c-Myc and thus the apoptosis of cells. In vivo experiments show that the topical application of c-Myc siRNAs delivered by SPACE-EGF through the skin can significantly inhibit the growth of melanoma tumors. This work may provide insight into the development of new transdermal drug carriers to treat a variety of skin disorders. PMID:27374619

  6. NAADP-Dependent Ca(2+) Signaling Controls Melanoma Progression, Metastatic Dissemination and Neoangiogenesis.

    Science.gov (United States)

    Favia, Annarita; Pafumi, Irene; Desideri, Marianna; Padula, Fabrizio; Montesano, Camilla; Passeri, Daniela; Nicoletti, Carmine; Orlandi, Augusto; Del Bufalo, Donatella; Sergi, Manuel; Ziparo, Elio; Palombi, Fioretta; Filippini, Antonio

    2016-01-01

    A novel transduction pathway for the powerful angiogenic factor VEGF has been recently shown in endothelial cells to operate through NAADP-controlled intracellular release of Ca(2+). In the present report the possible involvement of NAADP-controlled Ca(2+) signaling in tumor vascularization, growth and metastatic dissemination was investigated in a murine model of VEGF-secreting melanoma. Mice implanted with B16 melanoma cells were treated with NAADP inhibitor Ned-19 every second day for 4 weeks and tumor growth, vascularization and metastatization were evaluated. Control specimens developed well vascularized tumors and lung metastases, whereas in Ned-19-treated mice tumor growth and vascularization as well as lung metastases were strongly inhibited. In vitro experiments showed that Ned-19 treatment controls the growth of B16 cells in vitro, their migratory ability, adhesive properties and VEGFR2 expression, indicating NAADP involvement in intercellular autocrine signaling. To this regard, Ca(2+) imaging experiments showed that the response of B16 cells to VEGF stimulation is NAADP-dependent. The whole of these observations indicate that NAADP-controlled Ca(2+) signaling can be relevant not only for neoangiogenesis but also for direct control of tumor cells. PMID:26733361

  7. NAADP-Dependent Ca2+ Signaling Controls Melanoma Progression, Metastatic Dissemination and Neoangiogenesis

    Science.gov (United States)

    Favia, Annarita; Pafumi, Irene; Desideri, Marianna; Padula, Fabrizio; Montesano, Camilla; Passeri, Daniela; Nicoletti, Carmine; Orlandi, Augusto; Del Bufalo, Donatella; Sergi, Manuel; Ziparo, Elio; Palombi, Fioretta; Filippini, Antonio

    2016-01-01

    A novel transduction pathway for the powerful angiogenic factor VEGF has been recently shown in endothelial cells to operate through NAADP-controlled intracellular release of Ca2+. In the present report the possible involvement of NAADP-controlled Ca2+ signaling in tumor vascularization, growth and metastatic dissemination was investigated in a murine model of VEGF-secreting melanoma. Mice implanted with B16 melanoma cells were treated with NAADP inhibitor Ned-19 every second day for 4 weeks and tumor growth, vascularization and metastatization were evaluated. Control specimens developed well vascularized tumors and lung metastases, whereas in Ned-19-treated mice tumor growth and vascularization as well as lung metastases were strongly inhibited. In vitro experiments showed that Ned-19 treatment controls the growth of B16 cells in vitro, their migratory ability, adhesive properties and VEGFR2 expression, indicating NAADP involvement in intercellular autocrine signaling. To this regard, Ca2+ imaging experiments showed that the response of B16 cells to VEGF stimulation is NAADP-dependent. The whole of these observations indicate that NAADP-controlled Ca2+ signaling can be relevant not only for neoangiogenesis but also for direct control of tumor cells. PMID:26733361

  8. Immune-mediated regression of established B16F10 melanoma by intratumoral injection of attenuated Toxoplasma gondii protects against rechallenge

    OpenAIRE

    Baird, Jason R; Byrne, Katelyn T.; Lizotte, Patrick H; Toraya-Brown, Seiko; Scarlett, Uciane K.; Alexander, Matthew P.; Sheen, Mee Rie; Fox, Barbara A.; Bzik, David J.; Bosenberg, Marcus; Mullins, David W.; Turk, Mary Jo; Fiering, Steven

    2012-01-01

    Immune recognition of tumors can limit cancer development, but antitumor immune responses are often blocked by tumor-mediated immunosuppression. Since microbes or microbial constituents are powerful adjuvants to stimulate immune responses, we evaluated whether intratumoral administration of a highly immunogenic but attenuated parasite could induce rejection of an established poorly immunogenic tumor.

  9. Comparative Depigmentation Effects of Resveratrol and Its Two Methyl Analogues in α-Melanocyte Stimulating Hormone-Triggered B16/F10 Murine Melanoma Cells

    OpenAIRE

    Yoon, Hoon-Seok; Hyun, Chang-Gu; Lee, Nam-Ho; Park, Sung-Soo; Shin, Dong-Bum

    2016-01-01

    Previous research showed that resveratrol (trans-3,4′,5-trihydroxystilbene) and pinostilbene (trans-3-methoxy-4′,5-dihydroxystilbene) were able to inhibit tyrosinase directly; however, anti-melanogenic effects of pterostilbene (trans-3,5-dimethoxy-4′-hydroxystilbene) and resveratrol trimethyl ether (RTE) have not been compared. To investigate the hypopigmentation effects of pterostilbene and RTE, melanin contents and intracellular tyrosinase activity were determined by western blot analysis. ...

  10. Penurunan Aktivitas Tirosinase dan Jumlah Melanin oleh Fraksi Etil Asetat Buah Malaka (Phyllantus emblica) pada Mouse Melanoma B16 Cell-Line

    OpenAIRE

    Reti Hindritiani; Diah Dhianawaty; Muchtan Sujatno; Endang Sutedja; Setiawan

    2013-01-01

    Melanin accumulation can lead to hyperpigmentation, and if it occurs on the face can cause psychosocial problem. Depigmenting agents derived from plants are increasingly utilized. Agents being developed have to be effective in inhibiting melanin synthesis and should not be toxic to melanocyte. This study aimed was to examine the effect of ethyl acetate fraction from Phyllanthus emblica (P. emblica) fruit, also known as malaka fruit, towards melanine synthesis, which was measured from the mela...

  11. The use of Zymosan A and bacteria anchored to tumor cells for effective cancer immunotherapy: B16-F10 murine melanoma model.

    Science.gov (United States)

    Waldmannová, Eva; Caisová, Veronika; Fáberová, Julie; Sváčková, Petra; Kovářová, Markéta; Sváčková, Denisa; Kumžáková, Zuzana; Jačková, Adéla; Vácová, Nikol; Nedbalová, Pavla; Horká, Marie; Kopecký, Jan; Ženka, Jan

    2016-10-01

    The idea of using killed microorganisms or their parts for a stimulation of immunity in the cancer immunotherapy is very old, but the question of interactions and binding of these preparations to tumor cells has not been addressed so far. The attachment of Zymosan A and both Gram-positive and Gram-negative bacteria to tumor cells was tested in in vivo experiments. This binding was accomplished by charge interactions, anchoring based on hydrophobic chains and covalent bonds and proved to be crucial for a strong immunotherapeutic effect. The establishment of conditions for simultaneous stimulation of both Toll-like and phagocytic receptors led to very strong synergy. It resulted in tumor shrinkage and its temporary or permanent elimination. The role of neutrophils in cancer immunotherapy was demonstrated and the mechanism of their action (frustrated phagocytosis) was proposed. Finally, therapeutic approaches applicable for safe human cancer immunotherapy are discussed. Heat killed Mycobacterium tuberculosis covalently attached to tumor cells seems to be promising tool for this therapy.

  12. Down-regulation of tyrosinase, TRP-1, TRP-2 and MITF expressions by citrus press-cakes in murine B16 F10 melanoma

    Directory of Open Access Journals (Sweden)

    Sang Suk Kim

    2013-08-01

    Conclusions: Considering the anti–melanogenic activity and human safety, CCE is considered as a potential anti-melanogenic agent and may be effective for topical application for treating hyperpigmentation disorders.

  13. Down-regulation of tyrosinase, TRP-1, TRP-2 and MITF expressions by citrus press-cakes in murine B16 F10 melanoma

    Institute of Scientific and Technical Information of China (English)

    Sang Suk Kim; Min-Jin Kim; Young Hun Choi; Byung Kuk Kim; Kwang Sik Kim; Kyung Jin Park; Suk Man Park; Nam Ho Lee; Chang-Gu Hyun

    2013-01-01

    Objective: To investigate the suitability of citrus-press cakes, by-products of the juice industry as a source for the whitening agents for cosmetic industry. Methods:Ethylacetate extracts of citrus-press cakes (CCE) were examined for their anti-melanogenic potentials in terms of the inhibition of melanin production and mechanisim of melanogenesis by using Western Blot analysis with tyrosinese, tyrosinase-related protein-1 (TRP-1), TRP2, and microphthalmia-associated transcription factor (MITF) proteins. To apply the topical agents, citrus-press cakes was investigated the safety in human skin cell line. Finally flavonoid analysis of CCE was also determined by HPLC analysis. Results: Results indicated that CCE were shown to down-regulate melanin content in a dose-dependent pattern. The CCE inhibited tyrosinase, TRP-2, and MITF expressions in a dose-dependent manner. To test the applicability of CCE to human skin, we used MTT assay to assess the cytotoxic effects of CCE on human keratinocyte HaCaT cells. The CCE exhibited low cytotoxicity at 50 µg/mL. Characterization of the citrus-press cakes for flavonoid contents using HPLC showed varied quantity of rutin, narirutin, and hesperidin. Conclusions:Considering the anti-melanogenic activity and human safety, CCE is considered as a potential anti-melanogenic agent and may be effective for topical application for treating hyperpigmentation disorders.

  14. Drivers of melanoma susceptibility

    OpenAIRE

    Robles Espinoza, Carla Daniela

    2015-01-01

    Cutaneous melanoma is a cancer of melanocytes, the pigment-producing cells in our skin. It is one of the most aggressive human malignancies, constituting only about 2% of all dermatological cancers but being responsible for over 75% of all deaths from skin cancer. It has recently become a major public health problem, as it is now the fifth most common cancer in the United Kingdom after its incidence more than quadrupled in the last three decades. For these reasons, understanding the biologica...

  15. Adjuvant Therapy: Melanoma

    Directory of Open Access Journals (Sweden)

    Diwakar Davar

    2011-01-01

    Full Text Available With an incidence that is increasing at 2–5% per year, cutaneous melanoma is an international scourge that disproportionately targets young individuals. Despite much research, the treatment of advanced disease is still quite challenging. Immunotherapy with high-dose interferon-α2b or interleukin-2 benefits a select group of patients in the adjuvant and metastatic settings, respectively, with significant attendant toxicity. Advances in the biology of malignant melanoma and the role of immunomodulatory therapy have produced advances that have stunned the field. In this paper, we review the data for the use of interferon-α2b in various dosing ranges, vaccine therapy, and the role of radiotherapy in the adjuvant setting for malignant melanoma. Recent trials in the metastatic setting using anticytoxic T-lymphocyte antigen-4 (anti-CTLA-4 monoclonal antibody therapy and BRAF inhibitor therapy have demonstrated clear benefit with prolongation of survival. Trials investigating combinations of these novel agents with existing immunomodulators are at present underway.

  16. Principles of Melanoma Staging.

    Science.gov (United States)

    Boland, Genevieve M; Gershenwald, Jeffrey E

    2016-01-01

    Although now commonplace in contemporary cancer care, the systematic approach to classification of disease-specific cancers into a formalized staging system is a relatively modern concept. Overall, the goals of cancer staging are to characterize the status of cancer at a specific moment in time, risk stratify, facilitate prognostication, and inform clinical decision making. The revisions to the American Joint Committee on Cancer (AJCC) melanoma staging system over time reflect changes in our understanding of the biology of the disease. Since the 1st edition, where tumor thickness was defined anatomically by its relationship to the reticular or papillary dermis (Clark level) as well as tumor thickness (Breslow thickness), there have been significant strides in our use of clinicopathological variables to stratify low- versus high-risk patients. Management of the regional nodal basin has also changed dramatically over time, impacted by techniques such as lymphatic mapping and sentinel lymph node biopsy (SLNB) and changes in pathological evaluation of the regional lymph nodes. Additionally, stratification of distant metastases has evolved as survival outcomes have been shown to vary based upon anatomic site of metastases and serum lactate dehydrogenase levels. The variables in use in the current (7th edition) AJCC staging system are surrogate markers of biology with validated impact of survival outcomes. Going forward, it is likely that these and additional clinicopathological factors will be integrated with molecular and other correlates of melanoma tumor biology to further refine and personalize melanoma staging. PMID:26601861

  17. Surgery of Primary Melanomas

    International Nuclear Information System (INIS)

    Surgery remains the mainstay of melanoma therapy, regardless of the tumor site. Only the early diagnosis combined with proper surgical therapy currently gives patients affected by this malignancy the chance for a full cure. The main goal of surgical therapy is to provide the local control of the disease and to secure long-term survival of the patient without reasonable functional and esthetic impairment. The recommended method of biopsy—excisional biopsy, as an initial diagnostic and, to some extent, therapeutic procedure—is performed under local anesthesia as an elliptical incision with visual clear margins of 1–3 mm and with some mm of subcutaneous tissue. The extent of radical excision of the primary tumor (or scar after excisional biopsy) is based on the histopathologic characteristics of the primary tumor and usually consists of 1–2 cm margins with primary closure. The philosophy behind conducted randomized clinical trials has been to find the most conservative surgical approach that is able to guarantee the same results as more demolitive treatment. This has been the background of the trials designed to define the correct margins of excision around a primary cutaneous melanoma. Much less definition can be dedicated to the surgical management of patients with non-cutaneous melanomas

  18. A disguised Melanoma

    Directory of Open Access Journals (Sweden)

    Cláudia Sofia Rego

    2012-02-01

    Full Text Available Melanoma is a tumor that develops as a result of the malignant transformation of the melanocytes. There is a worldwide estimate of 132,000 new cases per year. This case study presents a 70-year-old male person with history of Diabetes Mellitus type 2 for 10 years and extensive psoriasis vulgaris for 6 years. The patient developed an ulcerated lesion in the plantar region of the right foot in one-year time period. The histological examination revealed an ulcerated malignant melanoma, Clark level V, 5.6 mm thick (Breslow. The lesion was surgically removed and the sentinel lymph node biopsy was negative. Initial conclusions revealed an advanced state of evolution of the primary tumor (TNM IIC. CAT scan detected gastric metastasis, reclassifying the illness as a TNM IV stage. Malignant melanoma may be difficult to diagnose, as it was possible to observe in this case study, where a foot ulcer was late diagnosed, delaying the diagnosis of a severe neoplasia with high mortality rate.

  19. Heart Transplant

    Science.gov (United States)

    ... 2015 A heart transplant gives a patient with congenital heart disease the opportunity to have a normal heart with ... pulmonary artery and left atrium. In patients with congenital heart disease, the surgeon may simultaneous transplant the lungs and ...

  20. Cytotoxicity evaluation of carbon-encapsulated iron nanoparticles in melanoma cells and dermal fibroblasts

    International Nuclear Information System (INIS)

    Carbon-encapsulated iron nanoparticles (CEINs) are emerging as promising biomedical tools due to their unique physicochemical properties. In this study, the cytotoxic effect of CEINs (the mean diameter distribution ranges 46–56 nm) has been explored by MTT, LDH leakage, Calcein-AM/propidium iodide (PI) and Annexin V-FITC/PI assays in human melanoma (HTB-140), mouse melanoma (B16-F10) cells, and human dermal fibroblasts (HDFs). The results demonstrated that CEINs produce mitochondrial and cell membrane cytotoxicities in a dose (0.0001–100 μg/ml)-dependent manner. Moreover, the studies elucidated some differences in cytotoxic effects between CEINs used as raw and purified materials composing of the carbon surface with acidic groups. Experiments showed that HTB-140 cells are more sensitive to prone early apoptotic events due to raw CEINs as compared to B16-F10 or HDF cells, respectively. Taken together, these results suggest that the amount of CEINs administered to cells and the composition of CEINs containing different amounts of iron as well as the carbon surface modification type is critical determinant of cytotoxic responses in both normal and cancer (melanoma) cells

  1. Antimetastatic Potentials of Dioscorea nipponica on Melanoma In Vitro and In Vivo

    Directory of Open Access Journals (Sweden)

    Mao-Lin Ho

    2011-01-01

    Full Text Available Recent studies have revealed pleiotropic anticancer and antiproliferative capabilities of Dioscorea nipponica Makino whereas the effect of this plant on metastasis of cancer cells has not been clearly clarified. In the present study, we extracted Dioscorea nipponica Makino with methanol (DNE1, chloroform (DNE2, ethyl acetate (DNE3, n-butanol (DNE4, and water (DNE5. We first demonstrate that DNE3 was found to be effective in reducing the lung metastases formation by about 99.5% as compared to vehicle-treated control animals. When a nontoxic concentration of the extract was treated directly to highly metastatic murin melanoma cells (B16F10 and human melanoma cells (A2058 in vitro, it exerted a dose-dependent inhibitory effect on the invasion (P<.001, motility (P<.001, secretion of MMPs (P<.001, and u-PA (P<.001 of both cell lines. To investigate the possible mechanisms involved in these events, we performed western blot analysis to find that DNE inhibited phosphorylation of Akt. A treatment with DNE3 to B16F10 cells also inhibited the activation of NF-κB and increased the expression of IkappaB. Taken together, these findings suggested that DNE3 could reduce the metastasis of melanoma cells, thereby constituting an adjuvant treatment for metastasis control.

  2. Magnetic and thermal behaviour of the amorphous ferromagnet Fe79B16Si5

    International Nuclear Information System (INIS)

    Spin waves in the amorphous ferromagnet Fe79B16Si5 are studied by Moessbauer effect spectroscopy. The magnetic hyperfine field (MHF) is measured at the Fe sites of such a ferromagnet, which exhibits a temperature dependence of the form, H(T)/H(0) = (1 - BT/sup 3/2/ - CT/sup 5/2/), indicative of spin wave excitations in amorphous ferromagnets. The T/sup 3/2/ behaviour and the distribution of the exchange interactions are studied in detail as a function of the MHF. The spin wave excitations constant B/sub 3/2/ = BT/sub C//sup 3/2/ = 0.3 +- 0.05 and C/sub 5/2/ = CT/sub C//sup 5/2/ = 0.3 +- 0.05,Are obtained by fitting the experimental data, and where T/sub C/ = 670 K. The results sh=ow that the contribution of C/sub 5/2/ is extremely effective above 124 K, while the ratio C/sub 5/2//C/sub 3/2/ = 1.0 indicates that the present magnetic interaction is of long range order character. On the other hand, fluctuations of the exchange interaction constant are found to decrease with increasing temperature. Some information regarding the directions of the magnetic moments are obtained during the study of the magnetic anisotropy course. The values of the Einstein and Debye temperatures as measured from the thermal shift results are theta/sub E/ = 250 K and theta/sub D/ = 350 K,And from the Moessbauer factor measurements theta/sub E/ = 165 K and theta/sub D/ = 285 K. (autho=r)

  3. Camphene isolated from essential oil of Piper cernuum (Piperaceae) induces intrinsic apoptosis in melanoma cells and displays antitumor activity in vivo.

    Science.gov (United States)

    Girola, Natalia; Figueiredo, Carlos R; Farias, Camyla F; Azevedo, Ricardo A; Ferreira, Adilson K; Teixeira, Sarah F; Capello, Tabata M; Martins, Euder G A; Matsuo, Alisson L; Travassos, Luiz R; Lago, João H G

    2015-11-27

    Natural monoterpenes were isolated from the essential oil of Piper cernuum Vell. (Piperaceae) leaves. The crude oil and the individual monoterpenes were tested for cytotoxicity in human tumor cell lineages and B16F10-Nex2 murine melanoma cells. In the present work we demonstrate the activity of camphene against different cancer cells, with its mechanism of action being investigated in vitro and in vivo in murine melanoma. Camphene induced apoptosis by the intrinsic pathway in melanoma cells mainly by causing endoplasmic reticulum (ER) stress, with release of Ca(2+) together with HmgB1 and calreticulin, loss of mitochondrial membrane potential and up regulation of caspase-3 activity. Importantly, camphene exerted antitumor activity in vivo by inhibiting subcutaneous tumor growth of highly aggressive melanoma cells in a syngeneic model, suggesting a promising role of this compound in cancer therapy. PMID:26471302

  4. Antitumor Activity of Kielmeyera Coriacea Leaf Constituents in Experimental Melanoma, Tested in Vitro and in Vivo in Syngeneic Mice

    Directory of Open Access Journals (Sweden)

    Carlos Rogério Figueiredo

    2014-10-01

    Full Text Available Purpose: The antitumor activity of Kielmeyera coriacea (Clusiaceae, a medicinal plant used in the treatment of parasitic, as well as fungal and bacterial infections by the Brazilian Cerrado population, was investigated. Methods: A chloroform extract (CE of K. coriacea was tested in the murine melanoma cell line (B16F10-Nex2 and a panel of human tumor cell lines. Tumor cell migration was determined by the wound-healing assay and the in vivo antitumor activity of CE was investigated in a melanoma cell metastatic model. 1H NMR and GC/MS were used to determine CE chemical composition. Results: We found that CE exhibited strong cytotoxic activity against murine melanoma cells and a panel of human tumor cell lines in vitro. CE also inhibited growth of B16F10-Nex2 cells at sub lethal concentrations, inducing cell cycle arrest at S phase, and inhibition of tumor cell migration. Most importantly, administration of CE significantly reduced the number of melanoma metastatic nodules in vivo. Chemical analysis of CE indicated the presence of the long chain fatty compounds, 1-eicosanol, 1-docosanol, and 2-nonadecanone as main constituents. Conclusion: These results indicate that K. coriacea is a promising medicinal plant in cancer therapy exhibiting antitumor activity both in vitro and in vivo against different tumor cell lines.

  5. High fat diet increases melanoma cell growth in the bone marrow by inducing osteopontin and interleukin 6

    Science.gov (United States)

    Chen, Guang-Liang; Luo, Yubin; Eriksson, Daniel; Meng, Xianyi; Qian, Cheng; Bäuerle, Tobias; Chen, Xiao-Xiang; Schett, Georg; Bozec, Aline

    2016-01-01

    The impact of metabolic stress induced by obesity on the bone marrow melanoma niche is largely unknown. Here we employed diet induced obese mice model, where mice received high-fat (HFD) or normal diet (ND) for 6 weeks before challenge with B16F10 melanoma cells. Tumor size, bone loss and osteoclasts numbers were assessed histologically in the tibial bones. For defining the molecular pathway, osteopontin knock-out mice, interleukin 6 neutralizing antibody or Janus kinase 2 inhibition were carried out in the same model. Mechanistic studies such as adipocyte-melanoma co-cultures for defining adipocyte induced changes of tumor cell proliferation and expression profiles were also performed. As results, HFD enhanced melanoma burden in bone by increasing tumor area and osteoclast numbers. This process was associated with higher numbers of bone marrow adipocytes expressing IL-6 in direct vicinity to tumor cells. Inhibition of IL-6 or of downstream JAK2 blocked HFD-induced tumor progression. Furthermore, the phenotypic changes of melanoma cells triggered macrophage and osteoclast accumulation accompanied by increased osteopontin expression. Osteopontin triggered osteoclastogenesis and also exerted a positive feedback loop to tumor cells, which was abrogated in its absence. Metabolic stress by HFD promotes melanoma growth in the bone marrow by an increase in bone marrow adipocytes and IL-6-JAK2-osteopontin mediated activation of tumor cells and osteoclast differentiation. PMID:27049717

  6. ADAM15 expression is downregulated in melanoma metastasis compared to primary melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Ungerer, Christopher; Doberstein, Kai [Pharmazentrum Frankfurt/ZAFES, University Hospital Goethe University Frankfurt, Frankfurt am Main (Germany); Buerger, Claudia; Hardt, Katja; Boehncke, Wolf-Henning [Department of Dermatology, Clinic of the Goethe-University, Theodor-Stern-Kai, Frankfurt (Germany); Boehm, Beate [Division of Rheumatology, Goethe University, Frankfurt am Main (Germany); Pfeilschifter, Josef [Pharmazentrum Frankfurt/ZAFES, University Hospital Goethe University Frankfurt, Frankfurt am Main (Germany); Dummer, Reinhard [Department of Pathology, Institute of Surgical Pathology, University Hospital, Zurich (Switzerland); Mihic-Probst, Daniela [Department of Dermatology, University Hospital Zurich (Switzerland); Gutwein, Paul, E-mail: p.gutwein@med.uni-frankfurt.de [Pharmazentrum Frankfurt/ZAFES, University Hospital Goethe University Frankfurt, Frankfurt am Main (Germany)

    2010-10-22

    Research highlights: {yields} Strong ADAM15 expression is found in normal melanocytes. {yields} ADAM15 expression is significantly downregulated in patients with melanoma metastasis. {yields} TGF-{beta} can downregulate ADAM15 expression in melanoma cells. {yields} Overexpression of ADAM15 in melanoma cells inhibits migration, proliferation and invasion of melanoma cells. {yields} Conclusion: ADAM15 represents an tumor suppressor protein in melanoma. -- Abstract: In a mouse melanoma metastasis model it has been recently shown that ADAM15 overexpression in melanoma cells significantly reduced the number of metastatic nodules on the lung. Unfortunately, the expression of ADAM15 in human melanoma tissue has not been determined so far. In our study, we characterized the expression of ADAM15 in tissue micro-arrays of patients with primary melanoma with melanoma metastasis. ADAM15 was expressed in melanocytes and endothelial cells of benign nevi and melanoma tissue. Importantly, ADAM15 was significantly downregulated in melanoma metastasis compared to primary melanoma. We further demonstrate that IFN-{gamma} and TGF-{beta} downregulate ADAM15 protein levels in melanoma cells. To investigate the role of ADAM15 in melanoma progression, we overexpressed ADAM15 in melanoma cells. Importantly, overexpression of ADAM15 in melanoma cells reduced the migration, invasion and the anchorage dependent and independent cell growth of melanoma cells. In summary, the downregulation of ADAM15 plays an important role in melanoma progression and ADAM15 act as a tumorsuppressor in melanoma.

  7. Boron nanoparticles inhibit turnour growth by boron neutron capture therapy in the murine B16-OVA model

    DEFF Research Database (Denmark)

    Petersen, Mikkel Steen; Petersen, Charlotte Christie; Agger, Ralf;

    2008-01-01

    Background: Boron neutron capture therapy usually relies on soluble, rather than particulate, boron compounds. This study evaluated the use of a novel boron nanoparticle for boron neutron capture therapy. Materials and Methods: Two hundred and fifty thousand B16-OVA tumour cells, pre...

  8. 75 FR 59071 - Airworthiness Directives; Bombardier, Inc. Model CL-600-2B16 (CL-604 Variants (Including CL-605...

    Science.gov (United States)

    2010-09-27

    ... in the Federal Register on June 2, 2010 (75 FR 30740). That NPRM proposed to correct an unsafe...'' under the DOT Regulatory Policies and Procedures (44 FR 11034, February 26, 1979); and 3. Will not have.... Model CL-600-2B16 (CL- 604 Variants (Including CL-605 Marketing Variant)) Airplanes AGENCY:...

  9. 75 FR 38011 - Airworthiness Directives; Bombardier, Inc. Model CL-600-2B16 (CL-604 Variant) Airplanes

    Science.gov (United States)

    2010-07-01

    ... products. That NPRM was published in the Federal Register on January 4, 2010 (75 FR 91). That NPRM proposed... Order 12866; 2. Is not a ``significant rule'' under the DOT Regulatory Policies and Procedures (44 FR.... Model CL-600-2B16 (CL- 604 Variant) Airplanes AGENCY: Federal Aviation Administration (FAA),...

  10. 75 FR 30740 - Airworthiness Directives; Bombardier, Inc. Model CL-600-2B16 (CL-604 Variants (Including CL-605...

    Science.gov (United States)

    2010-06-02

    ... Procedures (44 FR 11034, February 26, 1979); and 3. Will not have a significant economic impact, positive or... Directives; Bombardier, Inc. Model CL-600-2B16 (CL- 604 Variants (Including CL-605 Marketing Variant... addition to Bombardier Inc. Models CL-600-2B19, CL-600-2C10 and CL-600-2D24. The latter three models...

  11. Hispolon Decreases Melanin Production and Induces Apoptosis in Melanoma Cells through the Downregulation of Tyrosinase and Microphthalmia-Associated Transcription Factor (MITF Expressions and the Activation of Caspase-3, -8 and -9

    Directory of Open Access Journals (Sweden)

    Yi-Shyan Chen

    2014-01-01

    Full Text Available Hispolon is one of the most important functional compounds that forms Phellinus linteus (Berkeley & Curtis Teng. Hispolon has antioxidant, anti-inflammatory, antiproliferative and anticancer effects. In this study, we analyzed the functions of hispolon on melanogenesis and apoptosis in B16-F10 melanoma cells. The results demonstrated that hispolon is not an enzymatic inhibitor for tyrosinase; rather, it represses the expression of tyrosinase and the microphthalmia-associated transcription factor (MITF to reduce the production of melanin in α-melanocyte-stimulating hormone (α-MSH-stimulated B16-F10 cells at lower concentrations (less than 2 μM. In contrast, at higher concentration (greater than 10 μM, hispolon can induce activity of caspase-3, -8 and -9 to trigger apoptosis of B16-F10 cells but not of Detroit 551 normal fibroblast cells. Therefore, we suggest that hispolon has the potential to treat hyperpigmentation diseases and melanoma skin cancer in the future.

  12. Irradiation of choroidal melanoma with iodine 125 ophthalmic plaque

    International Nuclear Information System (INIS)

    Radioactive iodine 125 is a low-energy γ isotope with physical characteristics suitable for irradiation of intraocular tumors. Metal ophthalmic applicators have been designed to shield vital ocular structures while allowing irradiation of the tumor. We compared the radiation effects of iodine 125 and cobalt 60. The Greene melanoma was transplanted into the suprachoroidal space of rabbits. The tumor then grew as an intraocular mass, was irradiated, and was followed up for two months before enucleation. Histopathologic examiation defined the extent of the radiation damage to the tumor and other ocular structures from the iodine 125 and from the cobalt 60. The eye irradiated with iodine 125 suffered minimal radiation damage, whereas the tumor was sterilized. The eye irradiated with cobalt 60 showed substantial radiation damage, and the melanoma was incompletely treated. Our results support the use of iodine 125 in treating intraocular tumors. More research is needed as to optimum total dose and dose rate

  13. em cultura de células de melanoma

    Directory of Open Access Journals (Sweden)

    Fernanda Vieira Berti

    2007-01-01

    Full Text Available Identification, extraction, isolation and characterization of plant derived active molecules that modulate cell viability are of great clinical and pharmacological interest. Phytochemical tests have been used to demonstrate the presence of several compounds both from the primary and secondary metabolism of Aloe vera L. Aloin, an anthraquinone isolated from the chlorophyll parenchyma of A. vera L, is known for its biological activity. The antitumoral activity of aloin and the chlorophyll parenchyma extract was evaluated in vitro with the use of murine melanoma cell line B16F10. Cells incubated with 5% CO 2 at 37ºC were treated with different aloin and parenchyma extract concentrations. Both affected cellular stability, acting as modulators of tumoral cell viability. Experimental data were fitted to a developed mathematical model. From the fitting parameters we concluded that the chlorophyll parenchyma extract is 2.3 times more toxic to the cell than aloin.

  14. Isolation and Identification of Cancer Stem-Like Cells from Murine Melanoma Cell Lines

    Institute of Scientific and Technical Information of China (English)

    Jun Dou; Kai Hu; Ning Gu; Meng Pan; Ping Wen; Yating Li; Quan Tang; Lili Chu; Fengshu Zhao; Chuilian Jiang; Weihua Hu

    2007-01-01

    In current study, cancer stem-like cells in the murine melanoma B16F10 cells were investigated. CD phenotypes of the B16F10 cells were analyzed by flow cytometry, and the specific CD phenotype cells from the B16F10 cells were isolated by MACS. Then we used colony formation assay in soft agar media, the cell growth assay in serum-free culture media as well as the tumorigenicity investigation of the specific CD phenotype cells in C57BL/6 mice,respectively, to identify cancer stem-like cells in the B16F10 cells. The results showed that the B16F10 cells could form spherical clones in serum-free culture media, and the rate of clonegenesis of CD133+, CD44+ and CD44+CD133+ cells was higher than that of CD133-, CD44- and CD44+CD133- cells in soft agar media, respectively.The tumorigenic potential of CD133+, CD44+, CD44+CD133+ cells and CD44+CD133+CD24+ cells was stronger than that of CD133-, CD44-, CD44+CD133- cells and CD44+CD133+CD24- cells in mice, respectively. In conclusion, the CD44+CD133+CD24+ cells have some biological properties of cancer stem-like cells or are highly similar to the characteristics of cancer stem cells (CSC). These results provide an important method for identifying cancer stem-like cells in B16F10 cells and for further cancer target therapy.

  15. Characterization of post transplantation lymphoma in feline renal transplant recipients.

    Science.gov (United States)

    Durham, A C; Mariano, A D; Holmes, E S; Aronson, L

    2014-01-01

    The development of malignant neoplasia following solid organ transplantation and immunosuppression is well recognized in man. Post-transplantation malignant tumours include non-melanoma skin cancers, non-Hodgkin's lymphoma and Kaposi's sarcoma and many of these cancers have a known or suspected viral cause. A similar increased incidence of cancer is seen in cats that have received a renal transplant and lymphoma is the predominant neoplasm in this population. This study examines a population of cats that received renal transplants at the University of Pennsylvania School of Veterinary Medicine and subsequently developed neoplasia. From 1998 to 2010, 111 cats were transplanted and 25 cats developed cancer (22.5%). Fourteen of the 25 cats were diagnosed with lymphoma (56%), making it the most common tumour in this patient population. The median interval between transplantation and diagnosis of lymphoma was 617 days and the median survival time (MST) following the diagnosis of lymphoma was 2 days. Tissues from seven of these cats were available for histopathological review as either samples collected at necropsy examination (n = 5) or biopsy submissions (n = 2). Five of these cats had multiorgan involvement with sites including the liver, spleen, peripheral and mesenteric lymph nodes, small intestine, urinary bladder, heart, mesenteric fat and body wall. Four of the cats with multiorgan disease had involvement of the renal allograft two of which also had lymphoma of the native kidney. All lymphomas were classified as mid to high grade, diffuse large B-cell lymphoma, which is also the most common lymphoma subtype in human cases of post-transplantation lymphoproliferative disorders.

  16. Primary Desmoplastic Melanoma of the Penis

    OpenAIRE

    Chu, Julia T.; Liss, Michael A; Wu, William W.; Atreya Dash; Di Lu

    2015-01-01

    Desmoplastic melanomas are rare amelanotic melanomas that usually occur on skin with sun exposure. In this report, we present a 72-year-old man who presented with a desmoplastic melanoma of the penis. To our knowledge this represents the first reported case of primary desmoplastic melanoma of the penis. We discuss the pathologic differential and histologic evaluation.

  17. Photodynamic therapy for actinic keratosis in organ transplant patients

    DEFF Research Database (Denmark)

    Basset-Seguin, N; Baumann Conzett, K; Gerritsen, M J P;

    2013-01-01

    Background The incidence of actinic keratoses (AK) and non-melanoma skin cancer (NMSC) in organ transplant recipients (OTRs) is significantly higher than in immunocompetent patients. Rates of progression and recurrence following treatment are higher too, in part due to the effects...

  18. Effects of taurolidine on radiosensitivity of murine melanoma cells and its mechanism

    International Nuclear Information System (INIS)

    Objective: To observe the effects of taurolidine on radiosensitivity of B16-F10 cells of murine melanoma via the enhancement of Bax and Bad proteins and induction of Bcl-2 protein. Methods: The apoptosis of B16-F10 cells was assessed after treated with 0, 10, 25, 50, 100 and 150 μmol·L-1 taurolidine, clone survival assay was used to detect the radiosensitivity of B16-F10 cells, and protein expressions were determined by Western blotting. Results: The apoptosis of 5% cells was induced in a dose-and time-dependent manner after B16-F10 cells were treated with 50 μmol·L-1 taurolidine. The survival rate decreased after treated with tautolidine in combination with 2 Gy X-irradiation with the increase of taurolidine concentration and doses of irradiation (P0 and SER Dq) also increased with the increase of its concentration, there was significant difference between 50 μmol·L-1 taurolidine group and 10 μmol·L-1 taurolidine group (P<0.05); meantime, the level of proapototic protein Bax and Bad increased and the level of antiapoptotic protein Bcl-2 reduced. Conclusion: Taurolidine in combination with irradiation can enhance the radiosensitivity by the mediation of Bcl-2 family protein. (authors)

  19. A novel microtubule de-stabilizing complementarity-determining region C36L1 peptide displays antitumor activity against melanoma in vitro and in vivo

    Science.gov (United States)

    Figueiredo, Carlos R.; Matsuo, Alisson L.; Azevedo, Ricardo A.; Massaoka, Mariana H.; Girola, Natalia; Polonelli, Luciano; Travassos, Luiz R.

    2015-01-01

    Short peptide sequences from complementarity-determining regions (CDRs) of different immunoglobulins may exert anti-infective, immunomodulatory and antitumor activities regardless of the specificity of the original monoclonal antibody (mAb). In this sense, they resemble early molecules of innate immunity. C36L1 was identified as a bioactive light-chain CDR1 peptide by screening 19 conserved CDR sequences targeting murine B16F10-Nex2 melanoma. The 17-amino acid peptide is readily taken up by melanoma cells and acts on microtubules causing depolymerization, stress of the endoplasmic reticulum and intrinsic apoptosis. At low concentrations, C36L1 inhibited migration, invasion and proliferation of B16F10-Nex2 cells with cell cycle arrest at G2/M phase, by regulating the PI3K/Akt signaling axis involving Rho-GTPase and PTEN mediation. Peritumor injection of the peptide delayed growth of subcutaneously grafted melanoma cells. Intraperitoneal administration of C36L1 induced a significant immune-response dependent anti-tumor protection in a syngeneic metastatic melanoma model. Dendritic cells stimulated ex-vivo by the peptide and transferred to animals challenged with tumor cells were equally effective. The C36 VL CDR1 peptide is a promising microtubule-interacting drug that induces tumor cell death by apoptosis and inhibits metastases of highly aggressive melanoma cells. PMID:26391685

  20. Preventing Melanoma PSA (:60)

    Centers for Disease Control (CDC) Podcasts

    2015-06-02

    This 60 second public service announcement is based on the June 2015 CDC Vital Signs report. Skin cancer is the most common form of cancer in the U.S. In 2011, there were more than 65,000 cases of melanoma, the most deadly form of skin cancer. Learn how everyone can help prevent skin cancer.  Created: 6/2/2015 by National Center for Chronic Disease Prevention and Health Promotion (NCCDPHP).   Date Released: 6/2/2015.

  1. Lung transplantation

    Science.gov (United States)

    Afonso, José Eduardo; Werebe, Eduardo de Campos; Carraro, Rafael Medeiros; Teixeira, Ricardo Henrique de Oliveira Braga; Fernandes, Lucas Matos; Abdalla, Luis Gustavo; Samano, Marcos Naoyuki; Pêgo-Fernandes, Paulo Manuel

    2015-01-01

    ABSTRACT Lung transplantation is a globally accepted treatment for some advanced lung diseases, giving the recipients longer survival and better quality of life. Since the first transplant successfully performed in 1983, more than 40 thousand transplants have been performed worldwide. Of these, about seven hundred were in Brazil. However, survival of the transplant is less than desired, with a high mortality rate related to primary graft dysfunction, infection, and chronic graft dysfunction, particularly in the form of bronchiolitis obliterans syndrome. New technologies have been developed to improve the various stages of lung transplant. To increase the supply of lungs, ex vivo lung reconditioning has been used in some countries, including Brazil. For advanced life support in the perioperative period, extracorporeal membrane oxygenation and hemodynamic support equipment have been used as a bridge to transplant in critically ill patients on the waiting list, and to keep patients alive until resolution of the primary dysfunction after graft transplant. There are patients requiring lung transplant in Brazil who do not even come to the point of being referred to a transplant center because there are only seven such centers active in the country. It is urgent to create new centers capable of performing lung transplantation to provide patients with some advanced forms of lung disease a chance to live longer and with better quality of life. PMID:26154550

  2. Snake venoms components with antitumor activity in murine melanoma cells

    International Nuclear Information System (INIS)

    Despite the constant advances in the treatment of cancer, this disease remains one of the main causes of mortality worldwide. So, the development of new treatment modalities is imperative. Snake venom causes a variety of biological effects because they constitute a complex mixture of substances as disintegrins, proteases (serine and metalo), phospholipases A2, L-amino acid oxidases and others. The goal of the present work is to evaluate a anti-tumor activity of some snake venoms fractions. There are several studies of components derived from snake venoms with this kind of activity. After fractionation of snake venoms of the families Viperidae and Elapidae, the fractions were assayed towards murine melanoma cell line B16-F10 and fibroblasts L929. The results showed that the fractions of venom of the snake Notechis ater niger had higher specificity and potential antitumor activity on B16-F10 cell line than the other studied venoms. Since the components of this venom are not explored yet coupled with the potential activity showed in this work, we decided to choose this venom to develop further studies. The cytotoxic fractions were evaluated to identify and characterize the components that showed antitumoral activity. Western blot assays and zymography suggests that these proteins do not belong to the class of metallo and serine proteinases. (author)

  3. Ca2+-induced changes in energy metabolism and viability of melanoma cells

    OpenAIRE

    Glass-Marmor, L; Penso, J; Beitner, R

    1999-01-01

    Cancer cells are characterized by a high rate of glycolysis, which is their primary energy source. We show here that a rise in intracellular-free calcium ion (Ca2+), induced by Ca2+-ionophore A23187, exerted a deleterious effect on glycolysis and viability of B16 melanoma cells. Ca2+-ionophore caused a dose-dependent detachment of phosphofructokinase (EC 2.7.1.11), one of the key enzymes of glycolysis, from cytoskeleton. It also induced a decrease in the levels of glucose 1,6-bisphosphate and...

  4. Melanoma immunotherapy dominates the field.

    Science.gov (United States)

    Diamantopoulos, Panagiotis; Gogas, Helen

    2016-07-01

    The incidence of melanoma is increasing worldwide and despite early detection and intervention, the number of patients dying from metastatic disease continues to rise. The prognosis of advanced melanoma remains poor, with median survival between 6 and 9 months. Over the past 30 years and despite extensive clinical research, the treatment options for metastatic disease were limited and melanoma is still considered as one of the most therapy-resistant malignancies. Single-agent and combination chemotherapy, hormonal therapy, biochemotherapy, immunotherapy, targeted agent therapy and combination regimens failed to show a significant improvement in overall survival (OS). Recent advances and in-depth understanding of the biology of melanoma, have contributed to the development of new agents. Based on the molecular and immunological background of the disease, these new drugs have shown benefit in overall and progression-free survival (PFS). As the picture of the disease begins to change, oncologists need to alter their approach to melanoma treatment and consider disease biology together with targeted individualized treatment. In this review the authors attempt to offer an insight in the present and past melanoma treatment options, with a focus on the recently approved immunotherapeutic agents and the clinical perspectives of these new weapons against metastatic melanoma. PMID:27563656

  5. Human hemokinin-1 promotes migration of melanoma cells and increases MMP-2 and MT1-MMP expression by activating tumor cell NK1 receptors.

    Science.gov (United States)

    Zhang, Yixin; Li, Xiaofang; Li, Jingyi; Hu, Hui; Miao, Xiaokang; Song, Xiaoyun; Yang, Wenle; Zeng, Qian; Mou, Lingyun; Wang, Rui

    2016-09-01

    Receptors and their regulatory peptides are aberrantly expressed in tumors, suggesting a potential tumor therapy target. Human hemokinin-1 (hHK-1) is a tachykinin peptide ligand of the neurokinin-1 (NK1) receptor which is overexpressed in melanoma and other tumor tissues. Here, we investigated the role of hHK-1 and the NK1 receptor in melanoma cell migration. NK1 receptor expression was associated with melanoma metastatic potential. Treatment with hHK-1 significantly enhanced A375 and B16F10 melanoma cell migration and an NK1 receptor antagonist L732138 blocked this effect. MMP-2 and MT1-MMP expression were up-regulated in hHK-1-treated melanoma cells and cell signaling data suggested that hHK-1 induced phosphorylation of ERK1/2, JNK and p38 by way of PKC or PKA. Kinase activation led to increased MMP-2 and MT1-MMP expression and melanoma cell migration induced by hHK-1. Thus, hHK-1 and the NK1 receptor are critical to melanoma cell migration and each may be a promising chemotherapeutic target. PMID:27458061

  6. Melanoma Lentiginoso Acral

    Directory of Open Access Journals (Sweden)

    Gloria Andrea Vargas Suaza

    2008-12-01

    Full Text Available El melanoma lentiginoso acral (MLA es una variante rápidamente progresiva del melanoma maligno (MM. Constituye el 5-10% de todos los tipos de MM y se presenta con mayor frecuencia en pacientes de raza negra, asiáticos y latinoamericanos. En Colombia el MM se encuentra en aumento, con una incidencia de 3.5/100.000, siendo el MLA una de las variantes más comunes. La edad promedio de presentación es de 58 años, con una tasa de sobrevida menor para las personas de raza negra, asociado a un diagnóstico tardío. EL MLA se localiza en plantas, palmas y región subungueal y en su etiopatología se ha descrito la presencia de mutaciones en genes: 9p21 (p16: 67%, 11q13 (CCND1 (47%, 22q11-q13 (40% y 5p15 (20%. El diagnóstico de MLA, se ha fundamentado clásicamente en la histopatología. Herramientas de diagnóstico como la dermatoscopia, la evaluación del ganglio centinela y la determinación de alteraciones en las proteínas del ciclo celular contribuyen a la detección precoz del MLA y el MM en general.

  7. Studies of Secondary Melanoma on C57BL/6J Mouse Liver Using 1H NMR Metabolomics

    Energy Technology Data Exchange (ETDEWEB)

    Feng, Ju; Isern, Nancy G.; Burton, Sarah D.; Hu, Jian Z.

    2013-10-31

    NMR metabolomics, consisting of solid state high resolution (hr) magic angle spinning (MAS) 1H NMR (1H hr-MAS), liquid state high resolution 1H-NMR, and principal components analysis (PCA) has been used to study secondary metastatic B16-F10 melanoma in C57BL/6J mouse liver . The melanoma group can be differentiated from its control group by PCA analysis of the absolute concentrations or by the absolute peak intensities of metabolites from either 1H hr-MAS NMR data on intact liver tissues or liquid state 1H-NMR spectra on liver tissue extracts. In particular, we found that the absolute concentrations of alanine, glutamate, creatine, creatinine, fumarate and cholesterol are elevated in the melanoma group as compared to controls, while the absolute concentrations of succinate, glycine, glucose, and the family of linear lipids including long chain fatty acids, total choline and acylglycerol are decreased. The ratio of glycerophosphocholine to phosphocholine is increased by about 1.5 fold in the melanoma group, while the absolute concentration of total choline is actually lower in melanoma mice. These results suggest the following picture in secondary melanoma metastasis: Linear lipid levels are decreased by beta oxidation in the melanoma group, which contributes to an increase in the synthesis of cholesterol, and also provides an energy source input for TCA cycle. These findings suggest a link between lipid oxidation, the TCA cycle and the hypoxia-inducible factors (HIF) signal pathway in tumor metastases. Thus this study indicates that the metabolic profile derived from NMR analysis can provide a valuable bio-signature of malignancy and cell hypoxia in metastatic melanoma.

  8. A 99mTc(CO)3-labeled pyrazolyl-α-melanocyte-stimulating hormone analog conjugate for melanoma targeting

    International Nuclear Information System (INIS)

    Melanoma primary tumors can be, in most cases, removed surgically, whereas there is no satisfactory treatment for metastatic melanoma, being almost always lethal at this stage. Therefore, early detection of primary melanoma tumors is essential. The finding that melanocortin-1 receptor (MC1R) is overexpressed in isolated melanoma cells and melanoma tissues led to the radiolabeling of several α-melanocyte-stimulating hormone (α-MSH) analogs for early detection and treatment of melanoma. We have coupled the α-MSH analog Ac-Nle-Asp-His-D-Phe-Arg-Trp-Gly-Lys-NH2, through the ε-amino group of Lys11, to a pyrazolyl-containing chelator (pz). The resulting pz-α-MSH analog reacted with the fac-[99mTc(CO)3]+ moiety, giving [Ac-Nle4,Asp5,D-Phe7,Lys11(pz-99mTc(CO)3)]α-MSH4-11 in high yield, high specific activity and high radiochemical purity. This radioconjugate, which presents remarkable stability in vitro, exhibited time- and temperature-dependent internalization (4 h at 37oC; 56.7% maximum internalization) and high cellular retention (only 38% was released from the cell after 5 h) in murine melanoma B16F1 cells. A significant tumor uptake [4.2±0.9%ID/g, at 4 h postinjection (p.i.)] was also obtained in melanoma-bearing C57BL6 mice. The in vivo affinity and specificity of the radioconjugate to MC1R were demonstrated by receptor-blocking studies with the potent NDP-MSH agonist (63.5% reduction in tumor uptake at 4 h p.i.)

  9. Morphogenesis of early stage melanoma

    Science.gov (United States)

    Chatelain, Clément; Amar, Martine Ben

    2015-08-01

    Melanoma early detection is possible by simple skin examination and can insure a high survival probability when successful. However it requires efficient methods for identifying malignant lesions from common moles. This paper provides an overview first of the biological and physical mechanisms controlling melanoma early evolution, and then of the clinical tools available today for detecting melanoma in vivo at an early stage. It highlights the lack of diagnosis methods rationally linking macroscopic observables to the microscopic properties of the tissue, which define the malignancy of the tumor. The possible inputs of multiscale models for improving these methods are shortly discussed.

  10. Melanoma cell expression of CD200 inhibits tumor formation and lung metastasis via inhibition of myeloid cell functions.

    Directory of Open Access Journals (Sweden)

    Fatemeh Talebian

    Full Text Available CD200 is a cell surface glycoprotein that functions through engaging CD200 receptor on cells of the myeloid lineage and inhibits their functions. Expression of CD200 has been implicated in a variety of human cancer cells including melanoma cells and has been thought to play a protumor role. To investigate the role of cancer cell expression of CD200 in tumor formation and metastasis, we generated CD200-positive and CD200-negative B16 melanoma cells. Subcutaneous injection of CD200-positive B16 melanoma cells inhibited tumor formation and growth in C57BL/6 mice but not in Rag1⁻/⁻C57BL/6 mice. However, i.v. injection of CD200-positive B16 melanoma cells dramatically inhibited tumor foci formation in the lungs of both C57BL/6 and Rag1⁻/⁻C57BL6 mice. Flow cytometry analysis revealed higher expression of CD200R in Gr1⁺ myeloid cells in the lung than in peripheral myeloid cells. Depletion of Gr1⁺ cells or stimulation of CD200R with an agonistic antibody in vivo dramatically inhibited tumor foci formation in the lungs. In addition, treatment with tumor antigen specific CD4 or CD8 T cells or their combination yielded a survival advantage for CD200 positive tumor bearing mice over mice bearing CD200-negative tumors. Taken together, we have revealed a novel role for CD200-CD200R interaction in inhibiting tumor formation and metastasis. Targeting CD200R may represent a novel approach for cancer immunotherapy.

  11. Selective BRAF inhibition decreases tumor-resident lymphocyte frequencies in a mouse model of human melanoma.

    Science.gov (United States)

    Hooijkaas, Anna; Gadiot, Jules; Morrow, Michelle; Stewart, Ross; Schumacher, Ton; Blank, Christian U

    2012-08-01

    The development of targeted therapies and immunotherapies has markedly advanced the treatment of metastasized melanoma. While treatment with selective BRAF(V600E) inhibitors (like vemurafenib or dabrafenib) leads to high response rates but short response duration, CTLA-4 blocking therapies induce sustained responses, but only in a limited number of patients. The combination of these diametric treatment approaches may further improve survival, but pre-clinical data concerning this approach is limited. We investigated, using Tyr::CreER(T2)PTEN(F-/-)BRAF(F-V600E/+) inducible melanoma mice, whether BRAF(V600E) inhibition can synergize with anti-CTLA-4 mAb treatment, focusing on the interaction between the BRAF(V600E) inhibitor PLX4720 and the immune system. While PLX4720 treatment strongly decreased tumor growth, it did not induce cell death in BRAF(V600E)/PTEN(-/-) melanomas. More strikingly, PLX4720 treatment led to a decreased frequency of tumor-resident T cells, NK-cells, MDSCs and macrophages, which could not be restored by the addition of anti-CTLA-4 mAb. As this effect was not observed upon treatment of BRAF wild-type B16F10 tumors, we conclude that the decreased frequency of immune cells correlates to BRAF(V600E) inhibition in tumor cells and is not due to an off-target effect of PLX4720 on immune cells. Furthermore, anti-CTLA-4 mAb treatment of inducible melanoma mice treated with PLX4720 did not result in enhanced tumor control, while anti-CTLA-4 mAb treatment did improve the effect of tumor-vaccination in B16F10-inoculated mice. Our data suggest that vemurafenib may negatively affect the immune activity within the tumor. Therefore, the potential effect of targeted therapy on the tumor-microenvironment should be taken into consideration in the design of clinical trials combining targeted and immunotherapy.

  12. DOTA alpha-melanocyte-stimulating hormone analogues for imaging metastatic melanoma lesions.

    Science.gov (United States)

    Froidevaux, Sylvie; Calame-Christe, Martine; Sumanovski, Lazar; Tanner, Heidi; Eberle, Alex N

    2003-06-01

    Scintigraphic imaging of metastatic melanoma lesions requires highly tumor-specific radiopharmaceuticals. Because both melanotic and amelanotic melanomas overexpress melanocortin-1 receptors (MC1R), radiolabeled analogues of alpha-melanocyte-stimulating hormone (alpha-MSH) are potential candidates for melanoma diagnosis. Here, we report the in vivo performance of a newly designed octapeptide analogue, [betaAla(3), Nle(4), Asp(5), D-Phe(7), Lys(10)]-alpha-MSH(3-10) (MSH(OCT)), which was conjugated through its N-terminal amino group to the metal chelator 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) to enable incorporation of radiometals (e.g., indium-111) into the peptide. DOTA-MSH(OCT) displayed high in vitro MC1R affinity (IC(50) 9.21 nM). In vivo [(111)In]DOTA-MSH(OCT) exhibited a favorable biodistribution profile after injection in B16-F1 tumorbearing mice. The radiopeptide was rapidly cleared from blood through the kidneys and, most importantly, accumulated preferentially in the melanoma lesions. Lung and liver melanoma metastases could be clearly imaged on tissue section autoradiographs 4 h after injection of [(111)In]DOTA-MSH(OCT). A comparative study of [(111)In]DOTA-MSH(OCT) with [(111)In]DOTA-[Nle(4), D-Phe(7)]-alpha-MSH ([(111)In]-DOTA-NDP-MSH) demonstrated the superiority of the DOTA-MSH(OCT) peptide, particularly for the amount of radioactivity taken up by nonmalignant organs, including bone, the most radiosensitive tissue. These results demonstrate that [(111)In]DOTA-MSH(OCT) is a promising melanoma imaging agent.

  13. Melanoma Surveillance in the US: The Economic Burden of Melanoma

    Centers for Disease Control (CDC) Podcasts

    2011-10-19

    This podcast accompanies the publication of a series of articles on melanoma surveillance in the United States, available in the November supplement edition of the Journal of the American Academy of Dermatology. Dr. Gery Guy, from the CDC’s Division of Cancer Prevention and Control, discusses the economic burden of melanoma.  Created: 10/19/2011 by National Center for Chronic Disease Prevention and Health Promotion (NCCDPHP).   Date Released: 10/19/2011.

  14. Suppression of tumorigenicity and metastatic potential of melanoma cells by transduction of interferon gene

    Directory of Open Access Journals (Sweden)

    Lykhova A. A.

    2014-01-01

    Full Text Available The aim of this study was to investigate an inhibitory effect of baculovirus-mediated transduction of the murine interferon-beta gene on mouse melanoma in vitro and in vivo. Methods. Studies were performed on B16 mouse melanoma (MM-4 cell line. Transduction, immunocytochemical and tumor cell biology approaches have been used in this study. Results. Transduction of MM-4 cells by the recombinant baculovirus with IFN-beta gene is accompanied by morphological changes of tumor cells, suppression of cell proliferation, significant inhibition of platting efficiency of cells and their colonies formation in semisolid agar. Moreover, transduction of melanoma MM-4 cells by the baculovirus IFN-transgene leads to inhibition of tumorigenicity and metastatic ability of the cells in vivo. The intravenous administration of recombinant baculovirus vector with IFN gene inhibits growth of metastases induced in the lungs of mice by intravenously injected tumor cells. Conclusions. Transduction of mouse melanoma cells by the recombinant baculovirus with murine IFN-beta gene inhibits their proliferative potential, tumorigenicity and metastatic activity.

  15. Targeting DNA repair by coDbait enhances melanoma targeted radionuclide therapy.

    Science.gov (United States)

    Viallard, Claire; Chezal, Jean-Michel; Mishellany, Florence; Ranchon-Cole, Isabelle; Pereira, Bruno; Herbette, Aurélie; Besse, Sophie; Boudhraa, Zied; Jacquemot, Nathalie; Cayre, Anne; Miot-Noirault, Elisabeth; Sun, Jian-Sheng; Dutreix, Marie; Degoul, Françoise

    2016-03-15

    Radiolabelled melanin ligands offer an interesting strategy for the treatment of disseminated pigmented melanoma. One of these molecules, ICF01012 labelled with iodine 131, induced a significant slowing of melanoma growth. Here, we have explored the combination of [131I]ICF01012 with coDbait, a DNA repair inhibitor, to overcome melanoma radioresistance and increase targeted radionuclide therapy (TRT) efficacy. In human SK-Mel 3 melanoma xenograft, the addition of coDbait had a synergistic effect on tumor growth and median survival. The anti-tumor effect was additive in murine syngeneic B16Bl6 model whereas coDbait combination with [131I]ICF01012 did not increase TRT side effects in secondary pigmented tissues (e.g. hair follicles, eyes). Our results confirm that DNA lesions induced by TRT were not enhanced with coDbait association but, the presence of micronuclei and cell cycle blockade in tumor shows that coDbait acts by interrupting or delaying DNA repair. In this study, we demonstrate for the first time, the usefulness of DNA repair traps in the context of targeted radionuclide therapy. PMID:26887045

  16. Targeting DNA repair by coDbait enhances melanoma targeted radionuclide therapy

    Science.gov (United States)

    Viallard, Claire; Chezal, Jean-Michel; Mishellany, Florence; Ranchon-Cole, Isabelle; Pereira, Bruno; Herbette, Aurélie; Besse, Sophie; Boudhraa, Zied; Jacquemot, Nathalie; Cayre, Anne; Miot-Noirault, Elisabeth; Sun, Jian-Sheng; Dutreix, Marie; Degoul, Françoise

    2016-01-01

    Radiolabelled melanin ligands offer an interesting strategy for the treatment of disseminated pigmented melanoma. One of these molecules, ICF01012 labelled with iodine 131, induced a significant slowing of melanoma growth. Here, we have explored the combination of [131I]ICF01012 with coDbait, a DNA repair inhibitor, to overcome melanoma radioresistance and increase targeted radionuclide therapy (TRT) efficacy. In human SK-Mel 3 melanoma xenograft, the addition of coDbait had a synergistic effect on tumor growth and median survival. The anti-tumor effect was additive in murine syngeneic B16Bl6 model whereas coDbait combination with [131I]ICF01012 did not increase TRT side effects in secondary pigmented tissues (e.g. hair follicles, eyes). Our results confirm that DNA lesions induced by TRT were not enhanced with coDbait association but, the presence of micronuclei and cell cycle blockade in tumor shows that coDbait acts by interrupting or delaying DNA repair. In this study, we demonstrate for the first time, the usefulness of DNA repair traps in the context of targeted radionuclide therapy. PMID:26887045

  17. Construction of Egr-IFN γ and it's expression in B16 cells induced by ionizing irradiation

    International Nuclear Information System (INIS)

    Mouse IFN γ cDNA was ligated to downstream of Egr-1 promoter to construct Egr-IFN γ plasmid. The recombined plasmids were transfected into B16 cells with liposome, then the expression of IFN γ after different doses of X-ray irradiation and the time course of expression were detected by ELISA. The results showed that the expression levels in experimental groups were higher than that in the control group obviously (p<0.01); 6h after 2 Gy X-ray irradiation, the expression of IFN γ was highest (p<0.01); The B16 cells transfected were given 2 Gy X-ray irradiation every 24h, the expression level was highest after first irradiation (p<0.01) and then decrease gradually. The results indicated that ionizing irradiation can activate the Egr-IFN γ recombined plasmid and regulate the expression of IFN γ gene in B16 cells. The observation may be of potential significance in tumor therapy

  18. Kita driven expression of oncogenic HRAS leads to early onset and highly penetrant melanoma in zebrafish.

    Directory of Open Access Journals (Sweden)

    Cristina Santoriello

    Full Text Available BACKGROUND: Melanoma is the most aggressive and lethal form of skin cancer. Because of the increasing incidence and high lethality of melanoma, animal models for continuously observing melanoma formation and progression as well as for testing pharmacological agents are needed. METHODOLOGY AND PRINCIPAL FINDINGS: Using the combinatorial Gal4-UAS system, we have developed a zebrafish transgenic line that expresses oncogenic HRAS under the kita promoter. Already at 3 days transgenic kita-GFP-RAS larvae show a hyper-pigmentation phenotype as earliest evidence of abnormal melanocyte growth. By 2-4 weeks, masses of transformed melanocytes form in the tail stalk of the majority of kita-GFP-RAS transgenic fish. The adult tumors evident between 1-3 months of age faithfully reproduce the immunological, histological and molecular phenotypes of human melanoma, but on a condensed time-line. Furthermore, they show transplantability, dependence on mitfa expression and do not require additional mutations in tumor suppressors. In contrast to kita expressing melanocyte progenitors that efficiently develop melanoma, mitfa expressing progenitors in a second Gal4-driver line were 4 times less efficient in developing melanoma during the three months observation period. CONCLUSIONS AND SIGNIFICANCE: This indicates that zebrafish kita promoter is a powerful tool for driving oncogene expression in the right cells and at the right level to induce early onset melanoma in the presence of tumor suppressors. Thus our zebrafish model provides a link between kita expressing melanocyte progenitors and melanoma and offers the advantage of a larval phenotype suitable for large scale drug and genetic modifier screens.

  19. The effect of low-level laser irradiation (In-Ga-Al-AsP - 660 nm) on melanoma in vitro and in vivo

    International Nuclear Information System (INIS)

    It has been speculated that the biostimulatory effect of Low Level Laser Therapy could cause undesirable enhancement of tumor growth in neoplastic diseases. The aim of the present study is to analyze the behavior of melanoma cells (B16F10) in vitro and the in vivo development of melanoma in mice after laser irradiation. We performed a controlled in vitro study on B16F10 melanoma cells to investigate cell viability and cell cycle changes by the Tripan Blue, MTT and cell quest histogram tests at 24, 48 and 72 h post irradiation. The in vivo mouse model (male Balb C, n = 21) of melanoma was used to analyze tumor volume and histological characteristics. Laser irradiation was performed three times (once a day for three consecutive days) with a 660 nm 50 mW CW laser, beam spot size 2 mm2, irradiance 2.5 W/cm2 and irradiation times of 60s (dose 150 J/cm2) and 420s (dose 1050 J/cm2) respectively. There were no statistically significant differences between the in vitro groups, except for an increase in the hypodiploid melanoma cells (8.48 ± 1.40% and 4.26 ± 0.60%) at 72 h post-irradiation. This cancer-protective effect was not reproduced in the in vivo experiment where outcome measures for the 150 J/cm2 dose group were not significantly different from controls. For the 1050 J/cm2 dose group, there were significant increases in tumor volume, blood vessels and cell abnormalities compared to the other groups. LLLT Irradiation should be avoided over melanomas as the combination of high irradiance (2.5 W/cm2) and high dose (1050 J/cm2) significantly increases melanoma tumor growth in vivo

  20. The effect of low-level laser irradiation (In-Ga-Al-AsP - 660 nm on melanoma in vitro and in vivo

    Directory of Open Access Journals (Sweden)

    Bjordal Jan M

    2009-11-01

    Full Text Available Abstract Background It has been speculated that the biostimulatory effect of Low Level Laser Therapy could cause undesirable enhancement of tumor growth in neoplastic diseases. The aim of the present study is to analyze the behavior of melanoma cells (B16F10 in vitro and the in vivo development of melanoma in mice after laser irradiation. Methods We performed a controlled in vitro study on B16F10 melanoma cells to investigate cell viability and cell cycle changes by the Tripan Blue, MTT and cell quest histogram tests at 24, 48 and 72 h post irradiation. The in vivo mouse model (male Balb C, n = 21 of melanoma was used to analyze tumor volume and histological characteristics. Laser irradiation was performed three times (once a day for three consecutive days with a 660 nm 50 mW CW laser, beam spot size 2 mm2, irradiance 2.5 W/cm2 and irradiation times of 60s (dose 150 J/cm2 and 420s (dose 1050 J/cm2 respectively. Results There were no statistically significant differences between the in vitro groups, except for an increase in the hypodiploid melanoma cells (8.48 ± 1.40% and 4.26 ± 0.60% at 72 h post-irradiation. This cancer-protective effect was not reproduced in the in vivo experiment where outcome measures for the 150 J/cm2 dose group were not significantly different from controls. For the 1050 J/cm2 dose group, there were significant increases in tumor volume, blood vessels and cell abnormalities compared to the other groups. Conclusion LLLT Irradiation should be avoided over melanomas as the combination of high irradiance (2.5 W/cm2 and high dose (1050 J/cm2 significantly increases melanoma tumor growth in vivo.

  1. Hair Dyes Resorcinol and Lawsone Reduce Production of Melanin in Melanoma Cells by Tyrosinase Activity Inhibition and Decreasing Tyrosinase and Microphthalmia-Associated Transcription Factor (MITF) Expression

    OpenAIRE

    Shu-Mei Lee; Yi-Shyan Chen; Chih-Chien Lin; Kuan-Hung Chen

    2015-01-01

    Hair coloring products are one of the most important cosmetics for modern people; there are three major types of hair dyes, including the temporary, semi-permanent and permanent hair dyes. The selected hair dyes (such as ammonium persulfate, sodium persulfate, resorcinol and lawsone) are the important components for hair coloring products. Therefore, we analyzed the effects of these compounds on melanogenesis in B16-F10 melanoma cells. The results proved that hair dyes resorcinol and lawsone ...

  2. IL-24/MDA-7 Suppressed the Transplantation Tumor in BALB/c Mice

    Institute of Scientific and Technical Information of China (English)

    Jiancheng Xue; Changlin Wu; Yi Zhu; Lan Li; Xintang Dang; Shaoguang Qu; Fang Liu

    2006-01-01

    It is well-documented that interleukin-24 (IL-24) can induce apoptosis in a large spectrum of human cancer derived cell lines, but the effect of MDA-7/IL-24 gene transfer on mouse melanoma cells remains unknown. The eukaryotic expressing plasmid of IL-24 (pEGFP-IL-24) was constructed by DNA recombination technique. The recombination plasmid and empty vector were transfected into B16F0 cells and the expressions of IL-24 were determined by LSM, the proliferation of B16F0 cells was measured by MTT assay, and apoptosis rate and cell-cycle distribution of B16F0 cells were measured by FCM. The inhibitory effect of IL-24 gene transfection in mouse solid tumor was observed and measured. Compared with the control, the proliferation of B16F0 cells was inhibited by transfection with pEGFP-IL-24 and the G2/M phase of the transfected cells was also increased.Moreover, the percentage of mice with detectable tumor was decreased after inoculated with B16F0 cells transfected with pEGFP-IL-24. Growth rate of tumor in mouse model was significantly inhibited in IL-24 gene therapy group compared with the control. Proliferation of B16F0 cells was inhibited by pEGFP-IL-24 transfection.The intratumor injection of pEGFP-IL-24 could inhibit the growth of solid tumor in mice remarkably.

  3. [Choroidal melanoma - evolution and prognosis].

    Science.gov (United States)

    Chiruţa, Daria; Stan, Cristina

    2014-01-01

    Choroidal melanoma is the most common primary intraocular malignant tumor. We present the case of a 62 year old patient who was diagnosed with intraocular tumor in his right eye, for about three years. Regarding the fact that the patient refused any kind of treatment during this period, we just had the opportunity to monitor this case. Finally, the diagnosis was choroidal melanoma, confirmed by the histopathological exam.

  4. Perceived Intra-Family Melanoma Risk Communication

    OpenAIRE

    Loescher, Lois J.; Crist, Janice D.; Siaki, Leilani A.C.L.

    2009-01-01

    Melanoma is a skin cancer that can be deadly. Members of families with a strong history of melanoma have a high risk of melanoma occurrence or recurrence. Enhanced survival in these family members could be influenced by their knowledge of melanoma risk and by simple behaviors to decrease their risk or detect melanoma in its early, most curable, stage. Yet, there is minimal exploration on communication of risk or risk-modifying behaviors in melanoma at-risk families. In this study we described...

  5. Melanoma therapy: Check the checkpoints.

    Science.gov (United States)

    Furue, Masutaka; Kadono, Takafumi

    2016-02-01

    Recent mutational and translational studies have revealed that the Ras/Raf/mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) pathway plays a key role in melanomagenesis. Mutations in NRAS and BRAF are found in the majority of melanomas resulting in the formation of constitutively active NRAS and BRAF molecules, which leads to the proliferation and survival of melanoma cells through the activation of MEK/ERK signals. Inhibitors of BRAF or MEK significantly extend the progression-free survival and overall survival of melanoma patients compared with conventional chemotherapies. Combining BRAF and MEK inhibitors further enhances the clinical effectiveness. Cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) is an immune checkpoint molecule that downregulates T-cell activation by binding to B7 (CD80/CD86) molecules on antigen-presenting cells. Programmed death receptor ligand 1 on melanoma cells negatively regulates T-cell function by binding to the programmed death-1 (PD-1) receptor on T cells. Antibodies against CTLA-4 and PD-1 also enhance the survival of melanoma patients. In this review, we summarize the clinical effectiveness and adverse events of the BRAF inhibitors, MEK inhibitors and anti-immune checkpoint antibodies in melanoma treatment.

  6. Organ Transplantation

    Science.gov (United States)

    ... recipients to reduce the risk of transplant rejection. Rejection happens when your immune system attacks the new organ. If you have a transplant, you must take drugs the rest of your life to help keep your body from rejecting the new organ.

  7. Regulating Transplants

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Legislation to determine brain death is viewed as essential in controlling the organ transplant industry Organ transplant represents a very sensitive and complicated issue. Experts say the temporary administrative regulations recently promulgated by the Central Government are an important step, but relevant laws and regulations must follow. Among these, the

  8. Liver transplantation

    OpenAIRE

    Rodríguez-Perálvarez, M; De La Mata, M; Burroughs, A K

    2014-01-01

    Purpose of review: Long-term survival of liver transplant recipients is threatened by increased rates of de-novo malignancy and recurrence of hepatocellular carcinoma (HCC), both events tightly related to immunosuppression. Recent findings: There is accumulating evidence linking increased exposure to immunosuppressants and carcinogenesis, particularly concerning calcineurin inhibitors (CNIs), azathioprine and antilymphocyte agents. A recent study including 219 HCC transplanted patients sh...

  9. Transplant psychiatry.

    Science.gov (United States)

    Potts, S G

    2009-12-01

    Transplant units increasingly recognise a need for assistance from psychiatrists and psychologists in the assessment and management of potential transplant recipients and live donors. This arises from the various known associations between mental disorder and the need for transplantation; the intensifying requirement to select carefully among the potential recipients and donors of scarce human organs; and the drive to maximise transplant outcomes by optimising all aspects of treatment after surgery. There is good evidence that careful, protocol-guided selection among potential candidates for transplantation with alcoholic liver disease helps ensure outcomes at least as good as for other forms of liver disease. The evidence base in other areas is less robust, but the principles guiding the psychiatric assessment are broadly the same. There is an increasing need for psychiatric assessment of potential live organ donors, in order to minimise the risks they run, and in the case of altruistic donation this is now mandatory in UK law. PMID:21152475

  10. Tamoxifen inhibits tumor cell invasion and metastasis in mouse melanoma through suppression of PKC/MEK/ERK and PKC/PI3K/Akt pathways

    Energy Technology Data Exchange (ETDEWEB)

    Matsuoka, Hiroshi [Division of Pharmacotherapy, Kinki University School of Pharmacy, Kowakae, Higashi-Osaka 577-8502 (Japan); Department of Pharmacy, Nara Hospital, Kinki University School of Medicine, 1248-1 Ikoma, Nara 630-0293 (Japan); Tsubaki, Masanobu [Division of Pharmacotherapy, Kinki University School of Pharmacy, Kowakae, Higashi-Osaka 577-8502 (Japan); Yamazoe, Yuzuru [Department of Pharmacy, Kinki University Hospital, Osakasayama, Osaka 589-8511 (Japan); Ogaki, Mitsuhiko [Department of Pharmacy, Higahiosaka City General Hospital, Higashi-osaka, Osaka 578-8588 (Japan); Satou, Takao; Itoh, Tatsuki [Department of Pathology, Kinki University School of Medicine, Osakasayama, Osaka 589-8511 (Japan); Kusunoki, Takashi [Department of Otolaryngology, Juntendo University School of Medicine, Tokyo (Japan); Nishida, Shozo, E-mail: nishida@phar.kindai.ac.jp [Division of Pharmacotherapy, Kinki University School of Pharmacy, Kowakae, Higashi-Osaka 577-8502 (Japan)

    2009-07-15

    In melanoma, several signaling pathways are constitutively activated. Among these, the protein kinase C (PKC) signaling pathways are activated through multiple signal transduction molecules and appear to play major roles in melanoma progression. Recently, it has been reported that tamoxifen, an anti-estrogen reagent, inhibits PKC signaling in estrogen-negative and estrogen-independent cancer cell lines. Thus, we investigated whether tamoxifen inhibited tumor cell invasion and metastasis in mouse melanoma cell line B16BL6. Tamoxifen significantly inhibited lung metastasis, cell migration, and invasion at concentrations that did not show anti-proliferative effects on B16BL6 cells. Tamoxifen also inhibited the mRNA expressions and protein activities of matrix metalloproteinases (MMPs). Furthermore, tamoxifen suppressed phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2) and Akt through the inhibition of PKC{alpha} and PKC{delta} phosphorylation. However, other signal transduction factor, such as p38 mitogen-activated protein kinase (p38MAPK) was unaffected. The results indicate that tamoxifen suppresses the PKC/mitogen-activated protein kinase kinase (MEK)/ERK and PKC/phosphatidylinositol-3 kinase (PI3K)/Akt pathways, thereby inhibiting B16BL6 cell migration, invasion, and metastasis. Moreover, tamoxifen markedly inhibited not only developing but also clinically evident metastasis. These findings suggest that tamoxifen has potential clinical applications for the treatment of tumor cell metastasis.

  11. The association of N-palmitoylethanolamine with the FAAH inhibitor URB597 impairs melanoma growth through a supra-additive action

    International Nuclear Information System (INIS)

    The incidence of melanoma is considerably increasing worldwide. Frequent failing of classical treatments led to development of novel therapeutic strategies aiming at managing advanced forms of this skin cancer. Additionally, the implication of the endocannabinoid system in malignancy is actively investigated. We investigated the cytotoxicity of endocannabinoids and their hydrolysis inhibitors on the murine B16 melanoma cell line using a MTT test. Enzyme and receptor expression was measured by RT-PCR and enzymatic degradation of endocannabinoids using radiolabeled substrates. Cell death was assessed by Annexin-V/Propidium iodine staining. Tumors were induced in C57BL/6 mice by s.c. flank injection of B16 melanoma cells. Mice were injected i.p. for six days with vehicle or treatment, and tumor size was measured each day and weighted at the end of the treatment. Haematoxylin-Eosin staining and TUNEL assay were performed to quantify necrosis and apoptosis in the tumor and endocannabinoid levels were quantified by HPLC-MS. Tube formation assay and CD31 immunostaining were used to evaluate the antiangiogenic effects of the treatments. The N-arachidonoylethanolamine (anandamide, AEA), 2-arachidonoylglycerol and N- palmitoylethanolamine (PEA) reduced viability of B16 cells. The association of PEA with the fatty acid amide hydrolase (FAAH) inhibitor URB597 considerably reduced cell viability consequently to an inhibition of PEA hydrolysis and an increase of PEA levels. The increase of cell death observed with this combination of molecules was confirmed in vivo where only co-treatment with both PEA and URB597 led to decreased melanoma progression. The antiproliferative action of the treatment was associated with an elevation of PEA levels and larger necrotic regions in the tumor. This study suggests the interest of targeting the endocannabinoid system in the management of skin cancer and underlines the advantage of associating endocannabinoids with enzymatic hydrolysis

  12. The Complement C3a Receptor Contributes to Melanoma Tumorigenesis by Inhibiting Neutrophil and CD4+ T Cell Responses.

    Science.gov (United States)

    Nabizadeh, Jamileh A; Manthey, Helga D; Steyn, Frederik J; Chen, Weiyu; Widiapradja, Alexander; Md Akhir, Fazrena N; Boyle, Glen M; Taylor, Stephen M; Woodruff, Trent M; Rolfe, Barbara E

    2016-06-01

    The complement peptide C3a is a key component of the innate immune system and a major fragment produced following complement activation. We used a murine model of melanoma (B16-F0) to identify a hitherto unknown role for C3a-C3aR signaling in promoting tumor growth. The results show that the development and growth of B16-F0 melanomas is retarded in mice lacking C3aR, whereas growth of established melanomas can be arrested by C3aR antagonism. Flow cytometric analysis showed alterations in tumor-infiltrating leukocytes in the absence of C3aR. Specifically, neutrophils and CD4(+) T lymphocyte subpopulations were increased, whereas macrophages were reduced. The central role of neutrophils was confirmed by depletion experiments that reversed the tumor inhibitory effects observed in C3aR-deficient mice and returned tumor-infiltrating CD4(+) T cells to control levels. Analysis of the tumor microenvironment showed upregulation of inflammatory genes that may contribute to the enhanced antitumor response observed in C3aR-deficient mice. C3aR deficiency/inhibition was also protective in murine models of BRAF(V600E) mutant melanoma and colon and breast cancer, suggesting a tumor-promoting role for C3aR signaling in a range of tumor types. We propose that C3aR activation alters the tumor inflammatory milieu, thereby promoting tumor growth. Therapeutic inhibition of C3aR may therefore be an effective means to trigger an antitumor response in melanoma and other cancers. PMID:27183625

  13. Reproducibility of self-reported melanoma risk factors in melanoma patients

    NARCIS (Netherlands)

    Waal, A.C. de; Rossum, M.M. van; Kiemeney, L.A.L.M.; Aben, K.K.H.

    2014-01-01

    As melanoma researchers continue to investigate environmental and lifestyle-related risk factors, questionnaire data remain important. The reproducibility of a questionnaire on melanoma risk factors was investigated using a test-retest approach in 389 Dutch melanoma patients. In 2011, 389 melanoma p

  14. Aldesleukin and Pembrolizumab in Treating Patients With Stage III-IV Melanoma

    Science.gov (United States)

    2016-04-21

    Metastatic Melanoma; Stage III Mucosal Melanoma of the Head and Neck; Stage IIIA Skin Melanoma; Stage IIIB Skin Melanoma; Stage IIIC Skin Melanoma; Stage IV Skin Melanoma; Stage IVA Mucosal Melanoma of the Head and Neck; Stage IVB Mucosal Melanoma of the Head and Neck; Stage IVC Mucosal Melanoma of the Head and Neck

  15. Host immunity contributes to the anti-melanoma activity of BRAF inhibitors.

    Science.gov (United States)

    Knight, Deborah A; Ngiow, Shin Foong; Li, Ming; Parmenter, Tiffany; Mok, Stephen; Cass, Ashley; Haynes, Nicole M; Kinross, Kathryn; Yagita, Hideo; Koya, Richard C; Graeber, Thomas G; Ribas, Antoni; McArthur, Grant A; Smyth, Mark J

    2013-03-01

    The BRAF mutant, BRAF(V600E), is expressed in nearly half of melanomas, and oral BRAF inhibitors induce substantial tumor regression in patients with BRAF(V600E) metastatic melanoma. The inhibitors are believed to work primarily by inhibiting BRAF(V600E)-induced oncogenic MAPK signaling; however, some patients treated with BRAF inhibitors exhibit increased tumor immune infiltration, suggesting that a combination of BRAF inhibitors and immunotherapy may be beneficial. We used two relatively resistant variants of Braf(V600E)-driven mouse melanoma (SM1 and SM1WT1) and melanoma-prone mice to determine the role of host immunity in type I BRAF inhibitor PLX4720 antitumor activity. We found that PLX4720 treatment downregulated tumor Ccl2 gene expression and decreased tumor CCL2 expression in both Braf(V600E) mouse melanoma transplants and in de novo melanomas in a manner that was coincident with reduced tumor growth. While PLX4720 did not directly increase tumor immunogenicity, analysis of SM1 tumor-infiltrating leukocytes in PLX4720-treated mice demonstrated a robust increase in CD8(+) T/FoxP3(+)CD4(+) T cell ratio and NK cells. Combination therapy with PLX4720 and anti-CCL2 or agonistic anti-CD137 antibodies demonstrated significant antitumor activity in mouse transplant and de novo tumorigenesis models. These data elucidate a role for host CCR2 in the mechanism of action of type I BRAF inhibitors and support the therapeutic potential of combining BRAF inhibitors with immunotherapy.

  16. High accuracy of family history of melanoma in Danish melanoma cases

    DEFF Research Database (Denmark)

    Wadt, Karin A W; Drzewiecki, Krzysztof T; Gerdes, Anne-Marie

    2015-01-01

    The incidence of melanoma in Denmark has immensely increased over the last 10 years making Denmark a high risk country for melanoma. In the last two decades multiple public campaigns have sought to increase the awareness of melanoma. Family history of melanoma is a known major risk factor...... probands who reported 199 cases of melanoma in relatives, of which 135 cases where in first degree relatives. We confirmed the diagnosis of melanoma in 77% of all relatives, and in 83% of first degree relatives. In 181 probands we validated the negative family history of melanoma in 748 first degree...... reporting a negative family history of melanoma in first degree relatives this information is correct. In clinical practice we recommend that melanoma diagnosis in relatives should be verified if possible, but even unverified reported melanoma cases in relatives should be included in the indication...

  17. Isolated malignant melanoma metastasis to the pancreas

    DEFF Research Database (Denmark)

    Larsen, Anne K; Krag, Christen; Geertsen, Poul;

    2013-01-01

    SUMMARY: Malignant melanomas rarely develop isolated pancreatic metastases. We describe a unique patient who is still alive 22 years following an isolated pancreatic melanoma metastasis, and we review the sparse literature in the field....

  18. Molecular probes for malignant melanoma imaging.

    Science.gov (United States)

    Ren, Gang; Pan, Ying; Cheng, Zhen

    2010-09-01

    Malignant melanoma represents a serious public health problem and is a deadly disease when it is diagnosed at late stage. Though (18)F-fluorodeoxyglucose ((18)F-FDG) positron emission tomography (PET) has been widely used clinically for melanoma imaging, other approaches to specifically identify, characterize, monitor and guide therapeutics for malignant melanoma are still needed. Consequently, many probes targeting general molecular events including metabolism, angiogenesis, hypoxia and apoptosis in melanoma have been successfully developed. Furthermore, probes targeting melanoma associated targets such as melanocortin receptor 1 (MC1R), melanin, etc. have undergone active investigation and have demonstrated high melanoma specificity. In this review, these molecular probes targeting diverse melanoma biomarkers have been summarized. Some of them may eventually contribute to the improvement of personalized management of malignant melanoma. PMID:20497118

  19. Tool to Distinguish Moles from Melanoma

    Science.gov (United States)

    “Moles to Melanoma: Recognizing the ABCDE Features” presents photos that show changes in individual pigmented lesions over time, and describes the different appearances of moles, dysplastic nevi, and melanomas.

  20. Particularities of melanoma in Romania

    International Nuclear Information System (INIS)

    Full text: Malignant tumor from melanocytic origin, cutaneous melanoma is the most aggressive cutaneous cancer. The particularities of melanoma in Romania are: 1. the increasing number of cases in a geographic area dominated by the third cutaneous phototype; 2. the high level of T, represented by high levels of Breslow and Clark invasion markers; 3. the lack of a National Melanoma Register; 4. the lymph node sentinel biopsy technique introduction is the present focus; this allows an appropriate N affiliation; 5. the increasing of sanitary education in order to clinical and dermoscopical follow-up of atypical moles and the study of risk factors for melanoma; 6. the creation of a melanoma local guide. The survival prognosis could be improved by: a) the early detection in 'in situ' stage of melanoma and b) the correct TNM and/or EORTC affiliation of the case, in order to achieve an appropriate treatment. Sentinel lymph node detection and biopsy offers a new area in the evaluation of cancer spread by detecting micrometastases and performing a complete regional evaluation of the disease. The presence or absence of the metastases in the regional lymph nodes is one of the most important factors in the survival of patients with primary cutaneous melanoma. Early diagnosis of regional lymph node metastases enables the identification of those patients who are candidates for adjuvant therapy and early therapeutic lymph node dissection. At the same time, the sentinel node biopsy procedure represents a sensitive staging method. The paper presents some of gold standard diagnosis cases, dermoscopy and routine HE examination, selected from a total of 356 cases. (author)

  1. Effect of long-wave UV radiation on mouse melanoma: An in vitro and in vivo study

    International Nuclear Information System (INIS)

    doses of UVA irradiation. This enhancement of adhesiveness might lead to an increase in binding tumor cells to the endothelial lining of vasculature in various internal organs if occurring also in vivo. A further novel observation is that UVA induced both decline in the expression of E-cadherin adhesion molecule and increase in the expression of the N-cadherin adhesion molecule. In addition, a significant decline in homotypic melanoma-melanoma adhesion (clustering) was observed, which might result in the reduction of E-cadherin expression. It appears that UVA irradiation might reduce melanoma-melanoma interaction through decreasing the expression of E-cadherin and simultaneously enhance the adhesiveness of melanoma cells to endothelium, which in part could be mediated by N-cadherin expression. The aim of the in vivo animal study was to confirm the physiological significance of previously obtained in vitro results and to determine whether UVA radiation might increase melanoma metastasis in vivo. The use of C57BL/6 mice and syngeneic melanoma cell lines B16-F1 and B16-F10 showed that mice, which were i.v. injected with B16-F1 melanoma cells and thereafter exposed to UVA developed significantly more lung metastases when compared with the non-UVA-exposed group. To study the mechanism behind this phenomenon, the direct effect of UVA-induced lung colonization capacity was examined by the in vitro exposure of B16-F1 cells. Alternatively, the UVA-induced immunosuppression, which might be involved in increased melanoma metastasis, was measured by standard contact hypersensitivity assay (CHS). It appears that the UVA-induced increase of metastasis in vivo might be caused by a combination of UVA-induced systemic immunosuppression, and to the lesser extent, it might be caused by the increased adhesiveness of UVA irradiated melanoma cells. Finally, the UVA effect on gene expression in mouse melanoma was determined by a cDNA array, which revealed UVA-induced changes in the 9

  2. Effect of long-wave UV radiation on mouse melanoma: An in vitro and in vivo study

    Energy Technology Data Exchange (ETDEWEB)

    Pastila, R.

    2006-04-15

    doses of UVA irradiation. This enhancement of adhesiveness might lead to an increase in binding tumor cells to the endothelial lining of vasculature in various internal organs if occurring also in vivo. A further novel observation is that UVA induced both decline in the expression of E-cadherin adhesion molecule and increase in the expression of the N-cadherin adhesion molecule. In addition, a significant decline in homotypic melanoma-melanoma adhesion (clustering) was observed, which might result in the reduction of E-cadherin expression. It appears that UVA irradiation might reduce melanoma-melanoma interaction through decreasing the expression of E-cadherin and simultaneously enhance the adhesiveness of melanoma cells to endothelium, which in part could be mediated by N-cadherin expression. The aim of the in vivo animal study was to confirm the physiological significance of previously obtained in vitro results and to determine whether UVA radiation might increase melanoma metastasis in vivo. The use of C57BL/6 mice and syngeneic melanoma cell lines B16-F1 and B16-F10 showed that mice, which were i.v. injected with B16-F1 melanoma cells and thereafter exposed to UVA developed significantly more lung metastases when compared with the non-UVA-exposed group. To study the mechanism behind this phenomenon, the direct effect of UVA-induced lung colonization capacity was examined by the in vitro exposure of B16-F1 cells. Alternatively, the UVA-induced immunosuppression, which might be involved in increased melanoma metastasis, was measured by standard contact hypersensitivity assay (CHS). It appears that the UVA-induced increase of metastasis in vivo might be caused by a combination of UVA-induced systemic immunosuppression, and to the lesser extent, it might be caused by the increased adhesiveness of UVA irradiated melanoma cells. Finally, the UVA effect on gene expression in mouse melanoma was determined by a cDNA array, which revealed UVA-induced changes in the 9

  3. Preliminary biological evaluation of acridinic compounds for a targeted combined chemo and internal radionuclide therapy for melanoma

    International Nuclear Information System (INIS)

    The increasing incidence of melanoma and a lack of effective therapy on the disseminated form induces the development of selective tissue-targeted therapies. The aim of the present work was a targeting approach combining a bimodality therapy with the same compound exhibiting both chemo and internal radionuclide therapeutic properties. Benzamides are known to present a specific affinity for melanoma tissue. Former studies have shown that with aromatic and hetero-aromatic analogues of N-(2-diethylaminoethyl)- 4-iodo benzamide (B.Z.A.), the affinity for melanoma was maintained. In this context, new compounds have been designed and synthesized conjugating a cytotoxic hetero-aromatic moiety, an amino-alkyl amidic side chain for melanoma targeting and a radioiodine for internal radionuclide therapy. Acridinic derivatives known as cytotoxic DNA-intercalating agents have been chosen for this study. The cytotoxic activity of fifteen new compounds has been tested in vitro on a panel of cell lines and the I.C.50 values were determined. The three first selected compounds have been further evaluated: in vivo, on B 16 F0 melanoma bearing C 57 B.L.6 mice to determine the pharmacological kinetic and namely the tumoral affinity. Two compounds exhibited a high, specific and long lasting concentration in melanoma tumor giving them a kinetic profile favourable for an application to radionuclide therapy; in vitro, using the 'colony forming' test on melanoma cells, for a first approach of association of chemo toxicity and radiotoxicity. Assessed on the ability of cells to form colonies, the inhibition observed with the association for a same molecule of chemo toxic and radio toxic doses was quite exactly the sum of the two separate effects, a result providing a first validation of the radio chemotherapy concept; in vitro, by a preliminary determination of molecular mechanisms. Compared to parent compounds, results confirmed a maintain of DNA-intercalating properties. These first results

  4. Facial Transplantation.

    Science.gov (United States)

    Russo, Jack E; Genden, Eric M

    2016-08-01

    Reconstruction of severe facial deformities poses a unique surgical challenge: restoring the aesthetic form and function of the face. Facial transplantation has emerged over the last decade as an option for reconstruction of these defects in carefully selected patients. As the world experience with facial transplantation grows, debate remains regarding whether such a highly technical, resource-intensive procedure is warranted, all to improve quality of life but not necessarily prolong it. This article reviews the current state of facial transplantation with focus on the current controversies and challenges, with particular attention to issues of technique, immunology, and ethics. PMID:27400850

  5. Characterization and Inducing Melanoma Cell Apoptosis Activity of Mannosylerythritol Lipids-A Produced from Pseudozyma aphidis.

    Science.gov (United States)

    Fan, Linlin; Li, Hongji; Niu, Yongwu; Chen, Qihe

    2016-01-01

    Mannosylerythritol lipids (MELs) are natural glycolipid biosurfactants which have potential applications in the fields of food, cosmetic and medicine. In this study, MELs were produced from vegetable oil by Pseudozyma aphidis. Their structural data through LC/MS, GC/MS and NMR analysis revealed that MEL-A with two acetyls was the major compound and the identified homologs of MEL-A contained a length of C8 to C14 fatty acid chains. This glycolipid exhibited a surface tension of 27.69 mN/m at a critical micelle concentration (CMC), self-assembling into particles in the water solution. It was observed to induce cell growth-inhibition and apoptosis of B16 melanoma cells in a dose-dependent manner, as well as cause cell cycle arrest at the S phase. Further quantitative RT-PCR analysis and western blotting revealed an increasing tendency of both mRNA and protein expressions of Caspase-12, CHOP, GRP78 and Caspase-3, and a down-regulation of protein Bcl-2. Combined with the up regulation of signaling IRE1 and ATF6, it can be speculated that MEL-A-induced B16 melanoma cell apoptosis was associated with the endoplasmic reticulum stress (ERS). PMID:26828792

  6. Characterization and Inducing Melanoma Cell Apoptosis Activity of Mannosylerythritol Lipids-A Produced from Pseudozyma aphidis.

    Directory of Open Access Journals (Sweden)

    Linlin Fan

    Full Text Available Mannosylerythritol lipids (MELs are natural glycolipid biosurfactants which have potential applications in the fields of food, cosmetic and medicine. In this study, MELs were produced from vegetable oil by Pseudozyma aphidis. Their structural data through LC/MS, GC/MS and NMR analysis revealed that MEL-A with two acetyls was the major compound and the identified homologs of MEL-A contained a length of C8 to C14 fatty acid chains. This glycolipid exhibited a surface tension of 27.69 mN/m at a critical micelle concentration (CMC, self-assembling into particles in the water solution. It was observed to induce cell growth-inhibition and apoptosis of B16 melanoma cells in a dose-dependent manner, as well as cause cell cycle arrest at the S phase. Further quantitative RT-PCR analysis and western blotting revealed an increasing tendency of both mRNA and protein expressions of Caspase-12, CHOP, GRP78 and Caspase-3, and a down-regulation of protein Bcl-2. Combined with the up regulation of signaling IRE1 and ATF6, it can be speculated that MEL-A-induced B16 melanoma cell apoptosis was associated with the endoplasmic reticulum stress (ERS.

  7. In vitro incorporation of boronophenylalanine by amelanotic and melanotic murine and human malignant melanoma cell lines

    International Nuclear Information System (INIS)

    Pelleted cells were irradiated as previously described, following a 20 h incubation in RPM1 1640 medium, in the presence or absence of 10μg/ml D,L-paraboronophenylalanine hydrochloride (10B1-BPA.HCl). Thermal neutrons were derived from Moata, a 100-kW Argonaut-type light water reactor. The neutron flux was 2.6 x 109 n/cm2/s, dose rate 3.7 Gy/h (n+γ) and the dose range 0.6 - 0.8 Gy. Cells were plated onto X-irradiated feeder layers in triplicate in 25cm2 Falcon flasks and colonies counted after 11-12 days. No differences in thermal neutron radiosensitivity were observed for two amelanotic cell lines. A small but significant difference was observed for the melanotic cell line (418) grown in the presence or absence of BPA. Subsequent experiments showed that the uptake of boron was low in the B16 murine malignant melanoma cell line cultured in the presence of BPA. It was therefore necessary to investigate the boron uptake and incorporation in melanoma cells by increasing the BPA concentration, increasing melanization using different variants of the B16 cell lines, and alternative methods of cell layer detachment

  8. Coating Solid Lipid Nanoparticles with Hyaluronic Acid Enhances Antitumor Activity against Melanoma Stem-like Cells

    Science.gov (United States)

    Shen, Hongxin; Shi, Sanjun; Zhang, Zhirong; Gong, Tao; Sun, Xun

    2015-01-01

    Successful anticancer chemotherapy requires targeting tumors efficiently and further potential to eliminate cancer stem cell (CSC) subpopulations. Since CD44 is present on many types of CSCs, and it binds specially to hyaluronic acid (HA), we tested whether coating solid lipid nanoparticles with hyaluronan (HA-SLNs)would allow targeted delivery of paclitaxel (PTX) to CD44-overexpressing B16F10 melanoma cells. First, we developed a model system based on melanoma stem-like cells for experiments in vitro and in mouse xenografts, and we showed that cells expressing high levels of CD44 (CD44+) displayed a strong CSC phenotype while cells expressing low levels of CD44 (CD44-) did not. This phenotype included sphere and colony formation, higher proportion of side population cells, expression of CSC-related markers (ALDH, CD133, Oct-4) and tumorigenicity in vivo. Next we showed that administering PTX-loaded HA-SLNs led to efficient intracellular delivery of PTX and induced substantial apoptosis in CD44+ cells in vitro. In the B16F10-CD44+ lung metastasis model, PTX-loaded HA-SLNs targeted the tumor-bearing lung tissues well and subsequently exhibited significant antitumor effects with a relative low dose of PTX, which provided significant survival benefit without evidence of adverse events. These findings suggest that the HA-SLNs targeting system shows promise for enhancing cancer therapy. PMID:25897340

  9. Effects of Qingban Capsules on Melanin Synthesis of Murine B16 Cells%大豆异黄酮清斑软胶囊对小鼠B16细胞黑色素合成的影响

    Institute of Scientific and Technical Information of China (English)

    张国栋; 黄恺飞; 黄亦琦

    2012-01-01

    目的 测定大豆异黄酮清斑软胶囊对小鼠B16细胞黑色素合成的影响,研究其对黄褐斑的治疗作用. 方法 小鼠B16黑素细胞作为受试细胞,选择氢醌作为阳性对照物,用MTT比色法测定药物对细胞增殖的影响,采用酶学方法检测对酪氨酸酶活性的影响,NaOH溶解法测定黑素含量. 结果 不同浓度大豆异黄酮清斑软胶囊对黑素细胞增殖、酪氨酸酶活性及黑色素含量均有抑制作用,呈现剂量依赖性关系. 结论 大豆异黄酮清斑软胶囊能有效抑制黑素细胞增殖和降低酪氨酸酶活性,从而减少黑素合成.

  10. Ultra-structural cell distribution of the melanoma marker iodobenzamide: improved potentiality of SIMS imaging in life sciences

    Directory of Open Access Journals (Sweden)

    Papon Janine

    2004-04-01

    Full Text Available Abstract Background Analytical imaging by secondary ion mass spectrometry (SIMS provides images representative of the distribution of a specific ion within a sample surface. For the last fifteen years, concerted collaborative research to design a new ion microprobe with high technical standards in both mass and lateral resolution as well as in sensitivity has led to the CAMECA NanoSims 50, recently introduced onto the market. This instrument has decisive capabilities, which allow biological applications of SIMS microscopy at a level previously inaccessible. Its potential is illustrated here by the demonstration of the specific affinity of a melanoma marker for melanin. This finding is of great importance for the diagnosis and/or treatment of malignant melanoma, a tumour whose worldwide incidence is continuously growing. Methods The characteristics of the instrument are briefly described and an example of application is given. This example deals with the intracellular localization of an iodo-benzamide used as a diagnostic tool for the scintigraphic detection of melanic cells (e.g. metastasis of malignant melanoma. B16 melanoma cells were injected intravenously to C57BL6/J1/co mice. Multiple B16 melanoma colonies developed in the lungs of treated animals within three weeks. Iodobenzamide was injected intravenously in tumour bearing mice six hours before sacrifice. Small pieces of lung were prepared for SIMS analysis. Results Mouse lung B16 melanoma colonies were observed with high lateral resolution. Cyanide ions gave "histological" images of the cell, representative of the distribution of C and N containing molecules (e.g. proteins, nucleic acids, melanin, etc. while phosphorus ions are mainly produced by nucleic acids. Iodine was detected only in melanosomes, confirming the specific affinity of the drug for melanin. No drug was found in normal lung tissue. Conclusion This study demonstrates the potential of SIMS microscopy, which allows the

  11. Early detection and longitudinal monitoring of experimental primary and disseminated melanoma using [{sup 18}F]ICF01006, a highly promising melanoma PET tracer

    Energy Technology Data Exchange (ETDEWEB)

    Rbah-Vidal, Latifa; Vidal, Aurelien; Besse, Sophie; Audin, Laurent; Degoul, Francoise; Miot-Noirault, Elisabeth; Moins, Nicole; Auzeloux, Philippe; Chezal, Jean-Michel [Clermont Universite, Universite d' Auvergne, Imagerie Moleculaire et Therapie Vectorisee, BP 10448, Clermont-Ferrand (France); Inserm, U 990, Clermont-Ferrand (France); Cachin, Florent [Clermont Universite, Universite d' Auvergne, Imagerie Moleculaire et Therapie Vectorisee, BP 10448, Clermont-Ferrand (France); Inserm, U 990, Clermont-Ferrand (France); Centre Jean Perrin, Clermont-Ferrand (France); Bonnet, Mathilde [U1071 INSERM-Universite d' Auvergne, M2USH, Clermont-Ferrand (France); Askienazy, Serge [CYCLOPHARMA Laboratories, Biopole Clermont-Limagne, Saint-Beauzire (France); Dolle, Frederic [CEA, I2BM, Service Hospitalier Frederic Joliot, Orsay (France)

    2012-09-15

    Here, we report a new and rapid radiosynthesis of {sup 18}F-N-[2-(diethylamino)ethyl]-6-fluoro-pyridine-3-carboxamide ([{sup 18}F]ICF01006), a molecule with a high specificity for melanotic tissue, and its evaluation in a murine model for early specific detection of pigmented primary and disseminated melanoma. [{sup 18}F]ICF01006 was synthesized using a new one-step bromine-for-fluorine nucleophilic heteroaromatic substitution. Melanoma models were induced by subcutaneous (primary tumour) or intravenous (lung colonies) injection of B16BL6 melanoma cells in C57BL/6J mice. The relevance and sensitivity of positron emission tomography (PET) imaging using [{sup 18}F]ICF01006 were evaluated at different stages of tumoural growth and compared to {sup 18}F-fluorodeoxyglucose ([{sup 18}F]FDG). The fully automated radiosynthesis of [{sup 18}F]ICF01006 led to a radiochemical yield of 61 % and a radiochemical purity >99 % (specific activity 70-80 GBq/{mu}mol; total synthesis time 42 min). Tumours were visualized before they were palpable as early as 1 h post-injection with [{sup 18}F]ICF01006 tumoural uptake of 1.64 {+-} 0.57, 3.40 {+-} 1.47 and 11.44 {+-} 2.67 percentage of injected dose per gram of tissue (%ID/g) at days 3, 5 and 14, respectively. [{sup 18}F]ICF01006 PET imaging also allowed detection of melanoma pulmonary colonies from day 9 after tumour cell inoculation, with a lung radiotracer accumulation correlated with melanoma invasion. At day 21, radioactivity uptake in lungs reached a value of 5.23 {+-} 2.08 %ID/g (versus 0.41 {+-} 0.90 %ID/g in control mice). In the two models, comparison with [{sup 18}F]FDG showed that both radiotracers were able to detect melanoma lesions, but [{sup 18}F]ICF01006 was superior in terms of contrast and specificity. Our promising results provide further preclinical data, reinforcing the excellent potential of [{sup 18}F]ICF01006 PET imaging for early specific diagnosis and follow-up of melanin-positive disseminated melanoma. (orig.)

  12. Early detection and longitudinal monitoring of experimental primary and disseminated melanoma using [18F]ICF01006, a highly promising melanoma PET tracer

    International Nuclear Information System (INIS)

    Here, we report a new and rapid radiosynthesis of 18F-N-[2-(diethylamino)ethyl]-6-fluoro-pyridine-3-carboxamide ([18F]ICF01006), a molecule with a high specificity for melanotic tissue, and its evaluation in a murine model for early specific detection of pigmented primary and disseminated melanoma. [18F]ICF01006 was synthesized using a new one-step bromine-for-fluorine nucleophilic heteroaromatic substitution. Melanoma models were induced by subcutaneous (primary tumour) or intravenous (lung colonies) injection of B16BL6 melanoma cells in C57BL/6J mice. The relevance and sensitivity of positron emission tomography (PET) imaging using [18F]ICF01006 were evaluated at different stages of tumoural growth and compared to 18F-fluorodeoxyglucose ([18F]FDG). The fully automated radiosynthesis of [18F]ICF01006 led to a radiochemical yield of 61 % and a radiochemical purity >99 % (specific activity 70-80 GBq/μmol; total synthesis time 42 min). Tumours were visualized before they were palpable as early as 1 h post-injection with [18F]ICF01006 tumoural uptake of 1.64 ± 0.57, 3.40 ± 1.47 and 11.44 ± 2.67 percentage of injected dose per gram of tissue (%ID/g) at days 3, 5 and 14, respectively. [18F]ICF01006 PET imaging also allowed detection of melanoma pulmonary colonies from day 9 after tumour cell inoculation, with a lung radiotracer accumulation correlated with melanoma invasion. At day 21, radioactivity uptake in lungs reached a value of 5.23 ± 2.08 %ID/g (versus 0.41 ± 0.90 %ID/g in control mice). In the two models, comparison with [18F]FDG showed that both radiotracers were able to detect melanoma lesions, but [18F]ICF01006 was superior in terms of contrast and specificity. Our promising results provide further preclinical data, reinforcing the excellent potential of [18F]ICF01006 PET imaging for early specific diagnosis and follow-up of melanin-positive disseminated melanoma. (orig.)

  13. Surgical management of primary and recurrent melanoma.

    Science.gov (United States)

    Farma, Jeffrey M; Kulkarni, Nandini; Hsu, Cary

    2015-04-01

    Melanoma accounts for less than 2% of skin cancer cases but causes most skin cancer-related deaths. Surgery continues to be the cornerstone of treatment of melanoma and surgical principles are guided by data derived from clinical research. This article examines the evolution of surgical techniques for the diagnosis and treatment of primary and locally recurrent melanoma.

  14. Pathway aberrations of murine melanoma cells observed in Paired-End diTag transcriptomes

    Directory of Open Access Journals (Sweden)

    Liu Edison

    2007-06-01

    Full Text Available Abstract Background Melanoma is the major cause of skin cancer deaths and melanoma incidence doubles every 10 to 20 years. However, little is known about melanoma pathway aberrations. Here we applied the robust Gene Identification Signature Paired End diTag (GIS-PET approach to investigate the melanoma transcriptome and characterize the global pathway aberrations. Methods GIS-PET technology directly links 5' mRNA signatures with their corresponding 3' signatures to generate, and then concatenate, PETs for efficient sequencing. We annotated PETs to pathways of KEGG database and compared the murine B16F1 melanoma transcriptome with three non-melanoma murine transcriptomes (Melan-a2 melanocytes, E14 embryonic stem cells, and E17.5 embryo. Gene expression levels as represented by PET counts were compared across melanoma and melanocyte libraries to identify the most significantly altered pathways and investigate the expression levels of crucial cancer genes. Results Melanin biosynthesis genes were solely expressed in the cells of melanocytic origin, indicating the feasibility of using the PET approach for transcriptome comparison. The most significantly altered pathways were metabolic pathways, including upregulated pathways: purine metabolism, aminophosphonate metabolism, tyrosine metabolism, selenoamino acid metabolism, galactose utilization, nitrobenzene degradation, and bisphenol A degradation; and downregulated pathways: oxidative phosphorylation, ATPase synthesis, TCA cycle, pyruvate metabolism, and glutathione metabolism. The downregulated pathways concurrently indicated a slowdown of mitochondrial activities. Mitochondrial permeability was also significantly altered, as indicated by transcriptional activation of ATP/ADP, citrate/malate, Mg++, fatty acid and amino acid transporters, and transcriptional repression of zinc and metal ion transporters. Upregulation of cell cycle progression, MAPK, and PI3K/Akt pathways were more limited to certain

  15. JB/MS murine melanoma: a new model for studies on the modulation of differentiation and of tumorigenic and metastatic potential.

    Science.gov (United States)

    Hearing, V J; Cannon, G B; Vieira, W D; Jiménez-Atiénzar, M; Kameyama, K; Law, L W

    1988-02-15

    The recently obtained JB/MS melanoma (induced by DMBA in C57Bl/6 mice) has been successfully established in culture, and characterization of various parameters of these cells, as they have been serially passaged in vivo and in vitro, has begun. The culture lines were initially highly dendritic and melanotic, growing slowly in vitro and extremely slowly in vivo. During serial passage in vivo and in vitro the cell lines have gradually evolved into less melanotic, but more proliferative, tumorigenic and metastatic cells. We have been able to demonstrate that the JB/MS melanoma shares the common melanoma TSTA previously reported for B16, K1735 and JB/RH melanomas, but does not cross-react with the S91 melanoma or with other non-melanoma cell lines used as specificity controls. The JB/MS cells can be induced to differentiate in vitro by alpha-melanocyte stimulating hormone, a physiologically relevant agent, and studies have been initiated to detail the level at which this induction occurs. These sublines should prove to be excellent models for study of the progression of transformed cells from non-tumorigenic to tumorigenic phenotypes, and for progression through stages of varying metastatic potential, immunogenicity and differentiation.

  16. Melanoma stem cells in experimental melanoma are killed by radioimmunotherapy

    International Nuclear Information System (INIS)

    Introduction: In spite of recently approved B-RAF inhibitors and immunomodulating antibodies, metastatic melanoma has poor prognosis and novel treatments are needed. Melanoma stem cells (MSC) have been implicated in the resistance of this tumor to chemotherapy. Recently we demonstrated in a Phase I clinical trial in patients with metastatic melanoma that radioimmunotherapy (RIT) with 188-Rhenium(188Re)-6D2 antibody to melanin was a safe and effective modality. Here we investigated the interaction of MSC with RIT as a possible mechanism for RIT efficacy. Methods: Mice bearing A2058 melanoma xenografts were treated with either 1.5 mCi 188Re-6D2 antibody, saline, unlabeled 6D2 antibody or 188Re-labeled non-specific IgM. Results: On Day 28 post-treatment the tumor size in the RIT group was 4-times less than in controls (P < 0.001). The tumors were analyzed by immunohistochemistry and FACS for two MSC markers — chemoresistance mediator ABCB5 and H3K4 demethylase JARID1B. There were no significant differences between RIT and control groups in percentage of ABCB5 or JARID1B-positive cells in the tumor population. Our results demonstrate that unlike chemotherapy, which kills tumor cells but leaves behind MSC leading to recurrence, RIT kills MSC at the same rate as the rest of tumor cells. Conclusions: These results have two main implications for melanoma treatment and possibly other cancers. First, the susceptibility of ABCB5 + and JARID1B + cells to RIT in melanoma might be indicative of their susceptibility to antibody-targeted radiation in other cancers where they are present as well. Second, specifically targeting cancer stem cells with radiolabeled antibodies to ABCB5 or JARID1B might help to completely eradicate cancer stem cells in various cancers

  17. Liver Transplantation

    Science.gov (United States)

    ... to follow a treatment regimen a lack of psychosocial support In addition, the transplant candidate may decide ... is a challenging and complex surgery with significant risks to the donor, including death. According to the ...

  18. Lung transplant

    Science.gov (United States)

    Solid organ transplant - lung ... the new lung Have severe disease of other organs Cannot reliably take their medicines Are unable to ... medicines Damage to your kidneys, liver, or other organs from anti-rejection medicines Future risk of certain ...

  19. Kidney transplant

    Science.gov (United States)

    ... in cool salt water (saline) that preserves the organ for up to 48 hours. This gives the ... receive a transplanted kidney may reject the new organ. This means that their immune system sees the ...

  20. Pancreas Transplantation

    Science.gov (United States)

    The pancreas is a gland behind your stomach and in front of your spine. It produces the juices that ... hormones that help control blood sugar levels. A pancreas transplant is surgery to place a healthy pancreas ...

  1. Pancreas transplant

    Science.gov (United States)

    ... Heart tests such as an EKG, echocardiogram, or cardiac catheterization Tests to look for early cancer You will also want to consider one or more transplant centers to determine which is best for you: ...

  2. Corneal transplant

    Science.gov (United States)

    ... clear outer lens on the front of the eye. A corneal transplant is surgery to replace the cornea with tissue ... years. Rejection can sometimes be controlled with steroid eye drops. Other ... are: Bleeding Cataracts Infection of the eye Glaucoma ( ...

  3. Establishment of mouse CXCR3 gene-transfected cell line B16-mCXCR3 and study of its migration and oncogenic function in vitro and in vivo%转染小鼠CXCR3基因的B16-mCXCR3细胞在体内外迁移和致瘤作用研究

    Institute of Scientific and Technical Information of China (English)

    郭静雅; 朱华亭; 陈昌友; 黄莉; 刘玉华; 孙杰; 张彦军; 黄赛男; 邱玉华

    2011-01-01

    目的:建立稳定表达小鼠CXCR3基因的B16细胞株,研究CXCR3分子在肿瘤形成及转移过程中的作用.方法:采用PCR方法从pMD19-T/mCXCR3质粒中扩增CXCR3基因,插入到真核表达载体pIRES2-EGFP中,脂质体法转染小鼠黑色素瘤细胞B16;G418加压筛选阳性克隆,分别用RT-PCR方法与免疫荧光技术分析阳性克隆中CXCR3在mRNA和蛋白水平的表达.采用Transwell系统检测B16-mCXCR3细胞在其配体IP-10介导下的迁移能力.将B16-mCXCR3细胞分别通过皮下和眼静脉注射接种于BALB/c小鼠,观察在小鼠体内的成瘤率及肿瘤转移情况.结果:构建了表达小鼠CXCR3基因的真核表达载体pIRES2-EGFP/mCXCR3,转染该载体后获得了稳定表达CXCR3的B16-mCXCR3细胞.B16-mCXCR3细胞(1×105个),在20 ìg/L IP-10作用下迁移的细胞数为3 208个,与B16组和B16-mock组相比均具有统计学意义(P<0.01).于小鼠皮下接种B16-mCXCR3细胞、B16细胞和B16-mock细胞,第20天成瘤率均为100%.于小鼠眼静脉注射B16-mCXCR3细胞,第21天时50%的小鼠在肺部出现肉眼可见的黑色肿瘤转移灶,B16细胞组和B16-mock细胞组肺部未见肿瘤转移灶,B16-mCXCR3组与B16组和B16-mock组相比均具有统计学意义(P<0.01).结论:转染CXCR3基因的B16-mCXCR3细胞,在IP-10介导下可定向迁移,并可增加在小鼠体内的转移率.%Objective: In order to investigate the role of mouse gene CXCR3 in the process of tumor formation and metastasis, to construct an engineered B16 cell line expressing the mouse CXCR3 gene constantly.Methods: The eukaryotic transfer vector pIRES2-EGFP-mCXCB3 used to transfect B16 cells was constructed by inserting the PCR-amplified CXCR3 cassette from the plasmid pMD19-T/mCXCR3 between the EcoR I and BamH I sites of the eukaryotic expression vector pIRES2-EGFP and then the transfer vector was transfected into B16 cells by the liposome-mediated methods.The positive B16-mCXCR3 clones were screened for G418-resistance and

  4. Role of Acid Sphingomyelinase-Induced Signaling in Melanoma Cells for Hematogenous Tumor Metastasis

    Directory of Open Access Journals (Sweden)

    Alexander Carpinteiro

    2016-01-01

    Full Text Available Background: Hematogenous metastasis of malignant tumor cells is a multistep process that requires release of tumor cells from the local tumor mass, interaction of the tumor cells with platelets in the blood, and adhesion of either the activated tumor cells or the complexes of platelets and tumor cells to the endothelial cells of the target organ. We have previously shown that the interaction of melanoma cells with platelets results in the release of acid sphingomyelinase (Asm from activated platelets. Secreted platelet-derived Asm acts on malignant tumor cells to cluster and activate integrins; such clustering and activation are necessary for tumor cell adhesion to endothelial cells and for metastasis. Methods: We examined the response of tumor cells to treatment with extracellular sphingomyelinase or co-incubation with wild-type and Asm-deficient platelets. We determined the phosphorylation and activation of several intracellular signaling molecules, in particular p38 kinase (p38K, phospholipase Cγ (PLCγ, ezrin, and extracellular signal-regulated kinases. Results: Incubation of B16F10 melanoma cells with Asm activates p38 MAP kinase (p38K, phospholipase Cγ (PLCγ, ezrin, and extracellular signal-regulated kinases. Co-incubation of B16F10 melanoma cells with wild-type or Asm-deficient platelets showed that the phosphorylation/activation of p38K is dependent on Asm. Pharmacological blockade of p38K prevents activation of β1 integrin and adhesion in vitro. Most importantly, inhibition of p38K activity in B16F10 melanoma cells prevents tumor cell adhesion and metastasis to the lung in vivo, a finding indicating the importance of p38K for metastasis. Conclusions: Asm, secreted from activated platelets after tumor cell-platelet contact, induces p38K phosphorylation in tumor cells. This in turn stimulates β1 integrin activation that is necessary for adhesion and subsequent metastasis of tumor cells. Thus, inhibition of p38K might be a novel

  5. Familial skin cancer syndromes: Increased melanoma risk.

    Science.gov (United States)

    Ransohoff, Katherine J; Jaju, Prajakta D; Jaju, Prajaka D; Tang, Jean Y; Carbone, Michele; Leachman, Sancy; Sarin, Kavita Y

    2016-03-01

    Phenotypic traits, such as red hair and freckling, increase melanoma risk by 2- to 3-fold. In addition, approximately 10% of melanomas are caused by inherited germline mutations that increase melanoma risk from 4- to >1000-fold. This review highlights the key genes responsible for inherited melanoma, with an emphasis on when a patient should undergo genetic testing. Many genetic syndromes associated with increased melanoma risk are also associated with an increased risk of other cancers. Identification of these high-risk patients is essential for preventive behavior reinforcement, genetic counseling, and ensuring other required cancer screenings.

  6. Genetic alterations and markers of melanoma

    Directory of Open Access Journals (Sweden)

    N. N. Mazurenko

    2014-01-01

    Full Text Available Melanoma remains the most deadly form of malignant skin disease with high risk of metastases. Metastatic melanoma is prognostic highly unfavorable and resistant to traditional chemotherapy and biologic treatment. There is a great progress in understanding of the molecular mechanisms underlying melanoma initiation and progression. The external (ultraviolet irradiation and internal (genetic factors are involved in melanoma genesis. 5–14 % of melanoma cases occur in familial context due to genetic predisposition risk factors. Among them rare germinal mutations in the cell cycle genes regulators CDKN2A and CDK4 and in the master gene of melanocyte homeostasis MITF, as well as single nucleotide polymorphisms of several low-penetrated genes, namely MC1R, have been identified. The main cell signaling pathways and oncogene driver mutations are involved in melanoma pathogenesis. RAS / RAF / MEK / ERK cascade is hyperactivated in 75 % of cutaneous melanoma cases. Activation of PI3K / AKT / mTOR signaling pathway is important for melanoma progression. Recent studies revealed that melanomas are genetically and phenotypically heterogeneous tumors. Spectrum of chromosomal alterations and activating mutations corresponding to tumor molecular portraits varies in melanomas of different location. Most of cutaneous melanomas contain BRAF (50 % or NRAS (20 % mutations, and NRAS mutations occur on chronically sun-exposed skin. Activating KIT mutations have been reported in approximately 20–30 % of certain subtypes of melanoma, including acral and mucosal, and melanoma that develop on photodamaged skin. Cutaneous metastatic melanoma derive from preexisting nevi in 25 % of cases, molecular mechanisms of nevi malignization are discussed. Deepsequencing approaches of melanoma samples of different melanoma types highlighted new melanoma driver genes, that are damaged due to tumorigenic effects of ultraviolet: PPP6C, RAC1, SNX31, TACC1 and STK19. The

  7. Treatment of mouse melanoma cells with phorbol 12-myristate 13-acetate counteracts mannosylerythritol lipid-induced growth arrest and apoptosis.

    Science.gov (United States)

    Zhao, X; Geltinger, C; Kishikawa, S; Ohshima, K; Murata, T; Nomura, N; Nakahara, T; Yokoyama, K K

    2000-07-01

    Mannosylerythritol lipid (MEL), an extracellularglycolipid from yeast, induces the differentiation ofHL-60 promyelocytic leukemia cells towardsgranulocytes. We show here that MEL is also a potentinhibitor of the proliferation of mouse melanoma B16cells. Flow-cytometric analysis of the cell cycle ofMEL-treated B16 cells revealed the accumulation ofcells in the sub-G(0)/G(1) phase, which is a hallmark ofcells undergoing apoptosis. Treatment of B16 cellsfor 24 h with phorbol 12-myristate 13-acetate (PMA),an activator of protein kinase C (PKC), did notinterfere with the growth and survival of the cells,but it effectively counteracted the MEL-induced growtharrest and apoptosis. The activity of PKC was reducedin B16 cells treated with MEL at a concentration atwhich MEL induced apoptosis. However, incubation withPMA in addition to MEL reversed this reduction in theactivity of PKC. These results suggest thatconverging signaling pathways are triggeredindependently by MEL and PMA and that the signalsmight both be mediated by PKC. PMID:19002819

  8. International Transplant Nurses Society

    Science.gov (United States)

    ... 25th Annual ITNS Symposium The International Transplant Nurses Society (ITNS) cordially invites transplant nurses and other transplant ... pocket guide, developed by the International Transplant Nurses Society (ITNS), provides an overview of the interventions used ...

  9. Idiopathic Organ Transplant Chorioretinopathy after Liver Transplantation

    OpenAIRE

    Maria Fernanda Abalem; Pedro Carlos Carricondo; Sergio Luis Gianotti Pimentel; Walter Yukihiko Takahashi

    2015-01-01

    Idiopathic organ transplant chorioretinopathy is a rare disease associated with kidney and heart transplantation. We present a case secondary to liver transplantation including its multimodal imaging, differential diagnosis, and physiopathology discussion.

  10. Design of Pluronic-Based Formulation for Enhanced Redaporfin-Photodynamic Therapy against Pigmented Melanoma.

    Science.gov (United States)

    Pucelik, Barbara; Arnaut, Luis G; Stochel, Grażyna; Dąbrowski, Janusz M

    2016-08-31

    The therapeutic outcome of photodynamic therapy (PDT) with redaporfin (a fluorinated sulfonamide bacteriochlorin, F2BMet or LUZ11) was improved using Pluronic-based (P123, F127) formulations. Neither redaporfin encapsulated in Pluronic nor micelles alone exhibited cytotoxicity in a broad concentration range. Comprehensive in vitro studies against B16F10 melanoma cells showed that redaporfin-P123 micelles enhanced cellular uptake and increased oxidative stress compared with redaporfin-F127 or photosensitizer alone after short incubation times. ROS-sensitive fluorescent probes showed that the increased oxidative stress is due, at least in part, to a more efficient formation of hydroxyl radicals, and causes strong light-dose dependent apoptosis and necrosis. Tissue distribution and pharmacokinetic studies in tumor-bearing mice show that the Pluronic P123 formulation of redaporfin increases its bioavailability as well as the tumor-to-muscle and tumor-to-skin ratios, in comparison with Cremophor EL and Pluronic F127 formulations. Redaporfin in P123 was most successful in the PDT of C57BL/6J mice bearing subcutaneously implanted B16F10 melanoma tumors. Vascular-targeted PDT combining 1.5 mg kg(-1) redaporfin in P123 with a light dose of 74 J cm(-2) led to 100% complete cures (i.e., no tumor regrowth over one year post-treatment). This remarkable result reveals that modification of redaporfin with Pluronic block copolymers overcomes the resistance of melanoma cells to PDT possibly via increased tumor selectivity and enhanced ROS generation. PMID:27492026

  11. Protein engineering of insulin: Two novel fast-acting insulins [B16Ala]insulin and [B26Ala]insulin

    Institute of Scientific and Technical Information of China (English)

    ZHANG; Zhou; (张舟); TANG; Yuehua; (唐月华); YAO; Shiyin; (姚矢音); ZHU; Shangquan; (朱尚权); FENG; Youmin; (冯佑民)

    2003-01-01

    Blood glucose lowering assay proved that [B16Ala]insulin and [B26Ala]insulin exhibit potency of acute blood glucose lowering in normal pigs, which demonstrates that they are fast- acting insulin. Single-chain precursor of [B16Ala]insulin and [B26Ala]insulin is [B16Ala]PIP and [B26Ala]PIP, respectively, which are suitable for gene expression. Secretory expression level of the precursors in methylotrophic yeast Pichia pastoris was quite high, 650 mg/L and 130 mg/L, respectively. In vivo biological assay showed that the two fast-acting insulins have full or nearly full biological activity. So both [B16Ala]insulin and [B26Ala]insulin can be well developed as fast-acting insulin for clinic use.

  12. Resveratrol prevents inflammation-dependent hepatic melanoma metastasis by inhibiting the secretion and effects of interleukin-18

    Directory of Open Access Journals (Sweden)

    Valcarcel Maria

    2011-05-01

    Full Text Available Abstract Background Implantation and growth of metastatic cancer cells at distant organs is promoted by inflammation-dependent mechanisms. A hepatic melanoma metastasis model where a majority of metastases are generated via interleukin-18-dependent mechanisms was used to test whether anti-inflammatory properties of resveratrol can interfere with mechanisms of metastasis. Methods Two experimental treatment schedules were used: 1 Mice received one daily oral dose of 1 mg/kg resveratrol after cancer cell injection and the metastasis number and volume were determined on day 12. 2 Mice received one daily oral dose of 1 mg/kg resveratrol along the 5 days prior to the injection of cancer cells and both interleukin-18 (IL-18 concentration in the hepatic blood and microvascular retention of luciferase-transfected B16M cells were determined on the 18th hour. In vitro, primary cultured hepatic sinusoidal endothelial cells were treated with B16M-conditioned medium to mimic their in vivo activation by tumor-derived factors and the effect of resveratrol on IL-18 secretion, on vascular cell adhesion molecule-1 (VCAM-1 expression and on tumor cell adhesion were studied. The effect of resveratrol on melanoma cell activation by IL-18 was also studied. Results Resveratrol remarkably inhibited hepatic retention and metastatic growth of melanoma cells by 50% and 75%, respectively. The mechanism involved IL-18 blockade at three levels: First, resveratrol prevented IL-18 augmentation in the blood of melanoma cell-infiltrated livers. Second, resveratrol inhibited IL-18-dependent expression of VCAM-1 by tumor-activated hepatic sinusoidal endothelium, preventing melanoma cell adhesion to the microvasculature. Third, resveratrol inhibited adhesion- and proliferation-stimulating effects of IL-18 on metastatic melanoma cells through hydrogen peroxide-dependent nuclear factor-kappaB translocation blockade on these cells. Conclusions These results demonstrate multiple sites

  13. 76 FR 46597 - Airworthiness Directives; Bombardier, Inc. Model CL-600-2A12 (CL-601) and CL-600-2B16 (CL-601-3A...

    Science.gov (United States)

    2011-08-03

    .... Model CL-600-2A12 (CL- 601) and CL-600-2B16 (CL-601-3A, CL-601-3R, and CL-604 Variants) Airplanes... Bombardier, Inc. model-- Task(s)-- Initial compliance time (whichever occurs later)-- CL-600-2A12 (CL-601... icing. accumulation of 4,800 after the effective inclusive; and CL-600-2B16 (CL- total flight hours;...

  14. 76 FR 66203 - Airworthiness Directives; Bombardier, Inc. Model CL-600-2B16 (CL-601-3A, CL-601-3R, and CL-604...

    Science.gov (United States)

    2011-10-26

    ... Policies and Procedures (44 FR 11034, February 26, 1979); and 3. Will not have a significant economic.... Model CL-600-2B16 (CL- 601-3A, CL-601-3R, and CL-604 Variants) Airplanes AGENCY: Federal Aviation... Bombardier, Inc. Model CL-600-2B16 (CL- 601-3A, CL-601-3R, and CL-604 Variants) airplanes, certificated...

  15. Inhibitory Effects of Far-Infrared Irradiation Generated by Ceramic Material on Murine Melanoma Cell Growth

    Directory of Open Access Journals (Sweden)

    Ting-Kai Leung

    2012-01-01

    Full Text Available The biological effects of specific wavelengths, so-called “far-infrared radiation” produced from ceramic material (cFIR, on whole organisms are not yet well understood. In this study, we investigated the biological effects of cFIR on murine melanoma cells (B16-F10 at body temperature. cFIR irradiation treatment for 48 h resulted in an 11.8% decrease in the proliferation of melanoma cells relative to the control. Meanwhile, incubation of cells with cFIR for 48 h significantly resulted in 56.9% and 15.7% decreases in the intracellular heat shock protein (HSP70 and intracellular nitric oxide (iNO contents, respectively. Furthermore, cFIR treatment induced 6.4% and 12.3% increases in intracellular reactive oxygen species stained by 5-(and 6-carboxyl-2′,7′-dichlorodihydrofluorescein diacetate and dihydrorhodamine 123, respectively. Since malignant melanomas are known to have high HSP70 expression and iNO activity, the suppressive effects of cFIR on HSP70 and NO may warrant future interest in antitumor applications.

  16. YAG laser induced hyperthermia as an adjunctive treatment for ocular melanoma

    International Nuclear Information System (INIS)

    The correct treatment for choroidal melanoma is under debate, but radiation therapy (scleral plaques or external beam charged particles) is being used as the primary therapy in an increasing number of patients. Secondary effects of ocular radiation therapy are observed several years after treatment and can result in a significant decrease in visual acuity. Adjunctive use of ocular hyperthermia could reduce the dose of ionizing radiation needed to sterilize choroidal melanoma. In addition, if the production of ocular hyperthermia can be limited only to the tumor, then a reduction in the dose of ocular radiation could in turn reduce the severity of secondary complications to normal ocular structures. The authors performed preclinical studies with a low power continuous wave YAG laser as a source of precise and efficient hyperthermia. Tumor temperature profiles have been determined as a function of laser light power density and tumor function of laser light power density and tumor tissue depth in mice with B-16 melanomas. Ocular toxicity (in and out of the treatment field) induced by YAG laser hyperthermia has also been examined in pigmented rabbits using fundus photography, fluorrescein angiography and histopathology. Their results suggest that the YAG laser can generate efficient and highly localized hyperthermia, and may be useful in treating ocular tumors

  17. Semaphorin 3A suppresses tumor growth and metastasis in mice melanoma model.

    Directory of Open Access Journals (Sweden)

    Goutam Chakraborty

    Full Text Available BACKGROUND: Recent understanding on cancer therapy indicated that targeting metastatic signature or angiogenic switch could be a promising and rational approach to combat cancer. Advancement in cancer research has demonstrated the potential role of various tumor suppressor proteins in inhibition of cancer progression. Current studies have shown that axonal sprouting inhibitor, semaphorin 3A (Sema 3A acts as a potent suppressor of tumor angiogenesis in various cancer models. However, the function of Sema 3A in regulation of melanoma progression is not well studied, and yet to be the subject of intense investigation. METHODOLOGY/PRINCIPAL FINDINGS: In this study, using multiple in vitro and in vivo approaches we have demonstrated that Sema 3A acts as a potent tumor suppressor in vitro and in vivo mice (C57BL/6 models. Mouse melanoma (B16F10 cells overexpressed with Sema 3A resulted in significant inhibition of cell motility, invasiveness and proliferation as well as suppression of in vivo tumor growth, angiogenesis and metastasis in mice models. Moreover, we have observed that Sema 3A overexpressed melanoma clone showed increased sensitivity towards curcumin and Dacarbazine, anti-cancer agents. CONCLUSIONS: Our results demonstrate, at least in part, the functional approach underlying Sema 3A mediated inhibition of tumorigenesis and angiogenesis and a clear understanding of such a process may facilitate the development of novel therapeutic strategy for the treatment of cancer.

  18. Encapsulation of palladium porphyrin photosensitizer in layered metal oxide nanoparticles for photodynamic therapy against skin melanoma

    Science.gov (United States)

    Chen, Zih-An; Kuthati, Yaswanth; Kankala, Ranjith Kumar; Chang, Yu-Chuan; Liu, Chen-Lun; Weng, Ching-Feng; Mou, Chung-Yuan; Lee, Chia-Hung

    2015-10-01

    We designed a biodegradable nanocarrier of layered double hydroxide (LDH) for photodynamic therapy (PDT) based on the intercalation of a palladium porphyrin photosensitizer (PdTCPP) in the gallery of LDH for melanoma theragnosis. Physical and chemical characterizations have demonstrated the photosensitizer was stable in the layered structures. In addition, the synthesized nanocomposites rendered extremely efficacious therapy in the B16F10 melanoma cell line by improving the solubility of the hydrophobic PdTCPP photosensitizer. The detection of singlet oxygen generation under irradiation at the excitation wavelength of a 532 nm laser was indeed impressive. Furthermore, the in vivo results using a tumour xenograft model in mice indicated the apparent absence of body weight loss and relative organ weight variation to the liver and kidney demonstrated that the nanocomposites were biosafe with a significant reduction in tumour volume for the anti-cancer efficacy of PDT. This drug delivery system using the nanoparticle-photosensitizer hybrid has great potential in melanoma theragnosis.

  19. Tumor Heterogeneity in Uveal Melanomas

    NARCIS (Netherlands)

    H.W. Mensink (Hanneke)

    2010-01-01

    textabstractUveal melanoma (UM) is the most common primary intraocular malignancy in adults with an incidence of 7-10/ million and has a predilection for hematogenous dissemination to the liver. Despite improvements in diagnosis and treatment of this intraocular tumor, there has not been a change in

  20. Biology of Human Cutaneous Melanoma

    OpenAIRE

    Sharma, Bhuvnesh K.; Hasskamp, Joanne H.; Elias, Elias G.

    2010-01-01

    A review of the natural behavior of cutaneous melanoma, clinical and pathological factors, prognostic indicators, some basic research and the present and possible futuristic strategies in the management of this disease are presented. While surgery remains to be the most effective therapeutic approach in the management of early primary lesions, there is no standard adjuvant therapy after surgical resection, or for metastatic disease.

  1. [Malignant melanoma coexisting with pregnancy].

    Science.gov (United States)

    Krasomski, G; Broniarczyk, D; Gładysiak, A

    1992-09-01

    An extremely rare case of melanoma amelanoticum coexisting with pregnancy has been discussed. Pregnant A. Ch., age 42, was admitted to the Polish Mother's Health Centre Memorial Hospital on the 22nd of August, 1990 with a diagnosis of the 5th pregnancy, the 2nd delivery, the 30th week of gestation, state after cesarean section. Suspected malignant melanoma. Stomach ulceration. Thrombophlebitis of left lower extremity. General condition--medium hard. For the last three days she did not report fetal movements, fetal heartbeat was not detected either. Us examination confirmed fetal death. On the 24th of August, 1990, spontaneous vaginal delivery terminated the pregnancy, giving a dead, macerated female fetus, body weight of 1500 g. On the 3rd day after delivery the patient died with growing circulation-respiratory insufficiency. Autopsy revealed melanoma malignum amelanoticum disseminatum. Neither an autopsy of the fetus nor histopathological examinations of the secundines were performed for the advanced maceration. The coexistence of pregnancy with malignant melanoma in this case brought a tragic end both for the mother and the fetus. PMID:1305602

  2. Adjuvant therapy of malignant melanoma.

    Science.gov (United States)

    Molife, R; Hancock, B W

    2002-10-01

    High risk surgically resected melanoma is associated with a less than 50% 5-year survival. Adjuvant therapy is an appropriate treatment modality in this setting, and is more likely to be effective as the tumour burden here is small. Clinical observations of spontaneous tumour regressions and a highly variable rate of disease progression suggest a role of the immune system in the natural history of melanoma. Biological agents have therefore been the subjects of numerous adjuvant studies. Early, randomised controlled trials (RCTs) of Bacillus Calmette-Guerin (BCG), levamisole, Corynebacterium parvum, chemotherapy, isolated limb perfusion (ILP), radiotherapy, transfer factor (TF), megestrol acetate and vitamin A yielded largely negative results. Current trials focus on vaccines and the interferons. To date the latter is the only therapy to have shown a significant benefit in the prospective randomised controlled phase III setting. This report represents a systematic review of studies in adjuvant therapy in melanoma. Data from ongoing studies is awaited before a role for adjuvant agents in high risk melanoma is confirmed. PMID:12399001

  3. Prognostic stratification of ulcerated melanoma

    DEFF Research Database (Denmark)

    Bønnelykke-Behrndtz, Marie L; Schmidt, Henrik; Christensen, Ib J;

    2014-01-01

    OBJECTIVES: For patients with melanoma, ulceration is an important prognostic marker and interestingly also a predictive marker for the response of adjuvant interferon. A consensual definition and accurate assessment of ulceration are therefore crucial for proper staging and clinical management. We...

  4. Melanoma early detection and awareness

    DEFF Research Database (Denmark)

    Wainstein, Alberto; Algarra, Salvador Martin; Bastholt, Lars;

    2014-01-01

    Risk factors for melanoma are well known and have guided plans for primary and secondary prevention. The presentation of the disease, however, varies widely depending on the geographic area, ethnicity, and socioeconomic status. For this reason, many countries have developed specific strategies to...

  5. Pancreas transplants

    Energy Technology Data Exchange (ETDEWEB)

    Chandra, J.; Phillips, R.R.; Boardman, P.; Gleeson, F.V. [Department of Radiology, Churchill Hospital, Headington, Oxford (United Kingdom); Anderson, E.M. [Department of Radiology, Churchill Hospital, Headington, Oxford (United Kingdom)], E-mail: ewan.anderson@orh.nhs.uk

    2009-07-15

    Cadaveric, whole pancreas transplantation has proved an effective therapy in the treatment of long-standing type 1 diabetes mellitus and is capable of achieving an insulin-independent eugyclaemic state. As a result, this procedure is being increasingly performed. However, the surgical procedure is complex and unfamiliar to many radiologists. Imaging with computed tomography (CT) and magnetic resonance imaging (MRI) gives excellent results and can be used confidently to diagnose vascular, enteric, and immune-mediated complications. We present a review of the normal post-transplantation appearance and the features of early and late complications.

  6. Melanoma imaging using 111In-, 86Y- and 68Ga-labeled CHX-A''-Re(Arg11)CCMSH

    International Nuclear Information System (INIS)

    Introduction: A novel alpha-melanocyte-stimulating hormone peptide analog CHX-A''-Re(Arg11)CCMSH, which targeted the melanocortin-1 receptor (MC1-R) overexpressed on melanoma cells, was investigated for its biodistribution and tumor imaging properties. Methods: The metal bifunctional chelator CHX-A'' was conjugated to the melanoma targeting peptide (Arg11)CCMSH and cyclized by Re incorporation to yield CHX-A''-Re(Arg11)CCMSH. CHX-A''-Re(Arg11)CCMSH was labeled with 111In, 86Y and 68Ga, and the radiolabeled peptides were examined in B16/F1 melanoma-bearing mice for their pharmacokinetic as well as their tumor targeting properties using small animal SPECT and PET. Results: The radiolabeling efficiencies of the 111In-, 86Y- and 68Ga-labeled CHX-A''-Re(Arg11)CCMSH peptides were >95%, resulting in specific activities of 4.44, 3.7 and 1.85 MBq/μg, respectively. Tumor uptake of the 111In-, 86Y- and 68Ga-labeled peptides was rapid with 4.17±0.94, 4.68±1.02 and 2.68±0.69 %ID/g present in the tumors 2 h postinjection, respectively. Disappearance of radioactivity from the normal organs and tissues was rapid with the exception of the kidneys. Melanoma tumors were imaged with all three radiolabeled peptides 2 h postinjection. MC1-R-specific uptake was confirmed by competitive receptor blocking studies. Conclusions: Melanoma tumor uptake and imaging was exhibited by the 111In-, 86Y- and 68Ga-labeled Re(Arg11)CCMSH peptides, although the tumor uptake was moderated by low specific activity. The facile radiolabeling properties of CHX-A''-Re(Arg11)CCMSH allow it to be employed as a melanoma imaging agent with little or no purification after 111In, 86Y and 68Ga labeling.

  7. Novel photosensitizers trigger rapid death of malignant human cells and rodent tumor transplants via lipid photodamage and membrane permeabilization.

    Directory of Open Access Journals (Sweden)

    Mikhail M Moisenovich

    Full Text Available BACKGROUND: Apoptotic cascades may frequently be impaired in tumor cells; therefore, the approaches to circumvent these obstacles emerge as important therapeutic modalities. METHODOLOGY/PRINCIPAL FINDINGS: Our novel derivatives of chlorin e(6, that is, its amide (compound 2 and boronated amide (compound 5 evoked no dark toxicity and demonstrated a significantly higher photosensitizing efficacy than chlorin e(6 against transplanted aggressive tumors such as B16 melanoma and M-1 sarcoma. Compound 5 showed superior therapeutic potency. Illumination with red light of mammalian tumor cells loaded with 0.1 µM of 5 caused rapid (within the initial minutes necrosis as determined by propidium iodide staining. The laser confocal microscopy-assisted analysis of cell death revealed the following order of events: prior to illumination, 5 accumulated in Golgi cysternae, endoplasmic reticulum and in some (but not all lysosomes. In response to light, the reactive oxygen species burst was concomitant with the drop of mitochondrial transmembrane electric potential, the dramatic changes of mitochondrial shape and the loss of integrity of mitochondria and lysosomes. Within 3-4 min post illumination, the plasma membrane became permeable for propidium iodide. Compounds 2 and 5 were one order of magnitude more potent than chlorin e(6 in photodamage of artificial liposomes monitored in a dye release assay. The latter effect depended on the content of non-saturated lipids; in liposomes consisting of saturated lipids no photodamage was detectable. The increased therapeutic efficacy of 5 compared with 2 was attributed to a striking difference in the ability of these photosensitizers to permeate through hydrophobic membrane interior as evidenced by measurements of voltage jump-induced relaxation of transmembrane current on planar lipid bilayers. CONCLUSIONS/SIGNIFICANCE: The multimembrane photodestruction and cell necrosis induced by photoactivation of 2 and 5 are

  8. Kidney transplantation after liver transplantation

    Institute of Scientific and Technical Information of China (English)

    Li-Yang Wu; Hang Liu; Wei Liu; Han Li; Xiao-Dong Zhang

    2016-01-01

    Kidney transplantation after liver transplanta-tion (KALT) offers longer survival and a better quality of life to liver transplantation recipients who develop chronic renal failure. This article aimed to discuss the efifcacy and safety of KALT compared with other treatments. The medical records of 5 patients who had undergone KALT were retrospectively studied, together with a literature review of studies. Three of them developed chronic renal failure after liver transplanta-tion because of calcineurin inhibitor (CNI)-induced neph-rotoxicity, while the others had lupus nephritis or non-CNI drug-induced nephrotoxicity. No mortality was observed in the 5 patients. Three KALT cases showed good prognoses, maintaining a normal serum creatinine level during entire follow-up period. Chronic rejection occurred in the other two patients, and a kidney graft was removed from one of them. Our data suggested that KALT is a good alternative to dialysis for liver transplantation recipients. The cases also indicate that KALT can be performed with good long-term survival.

  9. The fibrinolytic system facilitates tumor cell migration across the blood-brain barrier in experimental melanoma brain metastasis

    International Nuclear Information System (INIS)

    Patients with metastatic tumors to the brain have a very poor prognosis. Increased metastatic potential has been associated with the fibrinolytic system. We investigated the role of the fibrinolytic enzyme plasmin in tumor cell migration across brain endothelial cells and growth of brain metastases in an experimental metastatic melanoma model. Metastatic tumors to the brain were established by direct injection into the striatum or by intracarotid injection of B16F10 mouse melanoma cells in C57Bl mice. The role of plasminogen in the ability of human melanoma cells to cross a human blood-brain barrier model was studied on a transwell system. Wild type mice treated with the plasmin inhibitor epsilon-aminocaproic acid (EACA) and plg-/- mice developed smaller tumors and survived longer than untreated wild type mice. Tumors metastasized to the brain of wild type mice treated with EACA and plg-/- less efficiently than in untreated wild type mice. No difference was observed in the tumor growth in any of the three groups of mice. Human melanoma cells were able to cross the human blood-brain barrier model in a plasmin dependent manner. Plasmin facilitates the development of tumor metastasis to the brain. Inhibition of the fibrinolytic system could be considered as means to prevent tumor metastasis to the brain

  10. Do We Know What Causes Melanoma Skin Cancer?

    Science.gov (United States)

    ... melanoma skin cancer be prevented? What causes melanoma skin cancer? Many risk factors for melanoma have been found, ... such as CDKN2A (also known as p16 ) and CDK4 that prevent them from doing their normal job ...

  11. Heart Transplantation

    Science.gov (United States)

    A heart transplant removes a damaged or diseased heart and replaces it with a healthy one. The healthy heart comes from a donor who has died. It is the last resort for people with heart failure when all other treatments have failed. The ...

  12. Melanoma biomarkers: Vox clamantis in deserto (Review).

    Science.gov (United States)

    Al-Shaer, Mays; Gollapudi, Divya; Papageorgio, Chris

    2010-05-01

    Detecting malignant melanoma at an early stage, monitoring therapy, predicting recurrence and identifying patients at risk for metastasis continue to be a challenging and demanding objective. The last two decades have witnessed innovations in the field of melanoma biomarkers. However, global agreement concerning monitoring and early detection has yet to be reached. This is a review of the current literature regarding melanoma biomarkers including demographic, clinical, pathological and molecular biomarkers that are produced by melanoma or non-melanoma cells. A number of these biomarkers demonstrate promising results as possible methods for early detection, predicting recurrence and monitoring therapy. Other biomarkers appear to be promising for identifying patients at risk for metastasis. We reviewed the most pertinent information in the field thus far and how this knowledge can impact, or not, the management of melanoma patients prognostically and therapeutically. PMID:22966315

  13. Melanoma-specific marker expression in skin biopsy tissues as a tool to facilitate melanoma diagnosis.

    Science.gov (United States)

    Alexandrescu, Doru T; Kauffman, C Lisa; Jatkoe, Timothy A; Hartmann, Dan P; Vener, Tatiana; Wang, Haiying; Derecho, Carlo; Rajpurohit, Yashoda; Wang, Yixin; Palma, John F

    2010-07-01

    Diagnosis of cutaneous melanoma requires accurate differentiation of true malignant tumors from highly atypical lesions, which lack the capacity to develop uncontrolled proliferation and to metastasize. We used melanoma markers from previous work to differentiate benign and atypical lesions from melanoma using paraffin-embedded tissue. This critical step in diagnosis generates the most uncertainty and discrepancy between dermatopathologists. A total of 193 biopsy tissues were selected: 47 melanomas, 48 benign nevi, and 98 atypical/suspicious, including 48 atypical nevi and 50 melanomas as later assigned by expert dermatopathologists. Performance for SILV, GDF15, and L1CAM normalized to TYR in unequivocal melanoma versus benign nevi resulted in an area under the curve (AUC) of 0.94, 0.67, and 0.5, respectively. SILV also differentiated atypical cases classified as melanoma from atypical nevi with an AUC=0.74. Furthermore, SILV showed a significant difference between suspicious melanoma and each suspicious atypia group: melanoma versus severe atypia and melanoma versus moderate atypia had P-values of 0.0077 and 0.0009, respectively. SILV showed clear discrimination between melanoma and benign unequivocal cases as well as between different atypia subgroups in the group of suspicious samples. The role and potential utility of this molecular assay as an adjunct to the morphological diagnosis of melanoma are discussed.

  14. Cu-64-labeled lactam bridge-cyclized α-MSH peptides for PET imaging of melanoma.

    Science.gov (United States)

    Guo, Haixun; Miao, Yubin

    2012-08-01

    The purpose of this study was to examine and compare the melanoma targeting and imaging properties of (64)Cu-NOTA-GGNle-CycMSH(hex) {(64)Cu-1,4,7-triazacyclononane-1,4,7-triacetic acid-Gly-Gly-Nle-c[Asp-His-DPhe-Arg-Trp-Lys]-CONH2} and (64)Cu-DOTA-GGNle-CycMSH(hex) {(64)Cu-1,4,7,10-tetraazacyclononane-1,4,7,10-tetraacetic acid-GGNle-CycMSH(hex)}. Two lactam bridge-cyclized peptides, NOTA-GGNle-CycMSH(hex) and DOTA-GGNle-CycMSH(hex), were synthesized using fluorenylmethyloxy carbonyl (Fmoc) chemistry. The melanocortin-1 (MC1) receptor binding affinity of NOTA-GGNle-CycMSH(hex) was determined in B16/F1 melanoma cells and compared with DOTA-GGNle-CycMSH(hex). The melanoma targeting and imaging properties of (64)Cu-NOTA-GGNle-CycMSH(hex) and (64)Cu-DOTA-GGNle-CycMSH(hex) were determined in B16/F1 melanoma-bearing C57 mice. NOTA-GGNle-CycMSH(hex) and DOTA-GGNle-CycMSH(hex) displayed comparable MC1 receptor binding affinities (1.6 vs 2.1 nM). The substitution of DOTA with NOTA dramatically increased the melanoma uptake and decreased the renal and liver uptake of (64)Cu-NOTA-GGNle-CycMSH(hex). The tumor uptake of (64)Cu-NOTA-GGNle-CycMSH(hex) was between 12.39 ± 1.61 and 12.71 ± 2.68% ID/g at 0.5, 2, and 4 h postinjection. The accumulation of (64)Cu-NOTA-GGNle-CycMSH(hex) activity in normal organs was lower than 1.02% ID/g except for the kidneys 2, 4, and 24 h postinjection. The tumor/liver uptake ratios of (64)Cu-NOTA-GGNle-CycMSHhex were 17.96, 16.95, and 8.02, whereas the tumor/kidney uptake ratios of (64)Cu-NOTA-GGNle-CycMSH(hex) were 2.52, 3.60, and 5.74 at 2, 4, and 24 h postinjection, respectively. Greater than 91% of the injected radioactivity cleared through the urinary system by 2 h postinjection. The substitution of DOTA with NOTA resulted in a dramatic increase in melanoma uptake and decrease in renal and liver uptake of (64)Cu-NOTA-GGNle-CycMSH(hex) as compared to (64)Cu-DOTA-GGNle-CycMSH(hex). High melanoma uptake coupled with low accumulation in nontarget

  15. Melanoma biomarkers: Vox clamantis in deserto (Review)

    OpenAIRE

    AL-SHAER, MAYS; GOLLAPUDI, DIVYA; PAPAGEORGIO, CHRIS

    2010-01-01

    Detecting malignant melanoma at an early stage, monitoring therapy, predicting recurrence and identifying patients at risk for metastasis continue to be a challenging and demanding objective. The last two decades have witnessed innovations in the field of melanoma biomarkers. However, global agreement concerning monitoring and early detection has yet to be reached. This is a review of the current literature regarding melanoma biomarkers including demographic, clinical, pathological and molecu...

  16. Melanoma of unknown origin: a case series.

    LENUS (Irish Health Repository)

    Kelly, J

    2010-12-01

    The natural history of metastatic melanoma involving lymph nodes, in the absence of a known primary site (cutaneous, ocular or mucosal) has, to date, been poorly defined; and the optimal management of this rare subtype of disease is therefore unclear. Melanomas of unknown primary site (MUP) are estimated to comprise between 3.7 and 6% of all melanomas (Anbari et al. in Cancer 79:1861-1821, 1997).

  17. Primary malignant melanoma of the esophagus

    International Nuclear Information System (INIS)

    Primary malignant melanoma most commonly originates from the skin; other less common extra cutaneous sites include squamous mucous membranes, uvea, retina, leptomeninges, genitourinary tract, digestive tract, biliary tract, and upper respiratory tract. Primary melanoma of the gastrointestinal tract is exceedingly rare. We are reporting a histo-pathologically proven rare case of primary malignant melanoma of the esophagus and its findings on fluorodeoxyglucose positron emission tomography and computed tomography

  18. Pediatric melanoma, moles, and sun safety.

    Science.gov (United States)

    Hawryluk, Elena B; Liang, Marilyn G

    2014-04-01

    Although pediatric melanoma is a rare disease, diagnosis and management of pigmented lesions in the pediatric population, particularly dysplastic nevi and Spitz nevi, can be challenging. In this article, we provide an overview of pigmented lesions in children, including melanoma and management of melanoma risk factors and melanocytic nevi in the pediatric population. Congenital melanocytic nevi, Spitz nevi, dysplastic and acquired nevi, and changes over time are reviewed. We discuss considerations for excision and management of pigmented lesions in children.

  19. Massive nodular melanoma scalp: a case report

    Directory of Open Access Journals (Sweden)

    A. Bhagya Lakshmi

    2015-04-01

    Full Text Available Melanoma is responsible for 1% to 2% of all cancer deaths around the world. Nodular melanoma often carries a poor prognosis because of no prodromal radial growth phase, early distant metastasis and significant tumour volume. We present a case of nodular melanoma measuring 20x10x8 cm in 28 year old tribal women. [Int J Res Med Sci 2015; 3(4.000: 1002-1005

  20. Primary rectal melanoma - a case report

    Directory of Open Access Journals (Sweden)

    Somak Das

    2015-01-01

    Full Text Available The most common site for malignant melanoma is skin, then eye and third is anorectal region. Primary anorectal malignant melanoma is still very uncommon. It is usually very aggressive and presents with altered bowel habit and rectal bleeding. Proctoscopy shows non-pigmented or lightly pigmented polypoid lesion. Histopathology is confirmatory. Early radical excision is mandatory. A 56 year-old female was presented with malignant melanoma of the lower third of rectum. We report this case for its rarity.

  1. Update in genetic susceptibility in melanoma

    OpenAIRE

    Potrony, Miriam; Badenas, Celia; Aguilera, Paula; Puig-Butille, Joan Anton; Carrera, Cristina; Malvehy, Josep; Puig, Susana

    2015-01-01

    Melanoma is the most deadly of the common skin cancers and its incidence is rapidly increasing. Approximately 10% of cases occur in a familial context. To date, cyclin-dependent kinase inhibitor 2A (CDKN2A), which was identified as the first melanoma susceptibility gene more than 20 years ago, is the main high-risk gene for melanoma. A few years later cyclin-dependent kinase 4 (CDK4) was also identified as a melanoma susceptibility gene. The technologic advances have allowed the identificatio...

  2. Histone variants and melanoma: facts and hypotheses.

    Science.gov (United States)

    Konstantinov, Nikifor K; Ulff-Møller, Constance J; Dimitrov, Stefan

    2016-07-01

    Melanoma is the most aggressive form of skin cancer with rising incidence and morbidity. Despite advances in treatment, the 10-yr survival for patients with metastatic disease is less than 10%. During the past few years, ongoing research on different epigenomic aberrations in melanoma has catalyzed better understanding of its pathogenesis and identification of new therapeutics. In our review, we will focus on the role of histone variants, key epigenetic players in melanoma initiation and progression. Specifically, incorporation of histone variants enables additional layers of chromatin structure, and here, we will describe how alterations in this epigenetic behavior impact melanoma.

  3. Metastatic Tumor Dormancy in Cutaneous Melanoma: Does Surgery Induce Escape?

    Directory of Open Access Journals (Sweden)

    William W. Tseng

    2011-02-01

    Full Text Available According to the concept of tumor dormancy, tumor cells may exist as single cells or microscopic clusters of cells that are clinically undetectable, but remain viable and have the potential for malignant outgrowth. At metastatic sites, escape from tumor dormancy under more favorable local microenvironmental conditions or through other, yet undefined stimuli, may account for distant recurrence after supposed “cure” following surgical treatment of the primary tumor. The vast majority of evidence to date in support of the concept of tumor dormancy originates from animal studies; however, extensive epidemiologic data from breast cancer strongly suggests that this process does occur in human disease. In this review, we aim to demonstrate that metastatic tumor dormancy does exist in cutaneous melanoma based on evidence from mouse models and clinical observations of late recurrence and occult transmission by organ transplantation. Experimental data underscores the critical role of impaired angiogenesis and immune regulation as major mechanisms for maintenance of tumor dormancy. Finally, we examine evidence for the role of surgery in promoting escape from tumor dormancy at metastatic sites in cutaneous melanoma.

  4. Metastatic Tumor Dormancy in Cutaneous Melanoma: Does Surgery Induce Escape?

    Energy Technology Data Exchange (ETDEWEB)

    Tseng, William W. [Department of Surgery, University of California at San Francisco, 513 Parnassus Avenue, Room S-321, San Francisco, CA 94143 (United States); Fadaki, Niloofar; Leong, Stanley P., E-mail: leongsx@cpmcri.org [Department of Surgery and Center for Melanoma Research and Treatment, California Pacific Medical Center and Research Institute, 2340 Clay Street, 2nd floor, San Francisco, CA 94115 (United States)

    2011-02-21

    According to the concept of tumor dormancy, tumor cells may exist as single cells or microscopic clusters of cells that are clinically undetectable, but remain viable and have the potential for malignant outgrowth. At metastatic sites, escape from tumor dormancy under more favorable local microenvironmental conditions or through other, yet undefined stimuli, may account for distant recurrence after supposed “cure” following surgical treatment of the primary tumor. The vast majority of evidence to date in support of the concept of tumor dormancy originates from animal studies; however, extensive epidemiologic data from breast cancer strongly suggests that this process does occur in human disease. In this review, we aim to demonstrate that metastatic tumor dormancy does exist in cutaneous melanoma based on evidence from mouse models and clinical observations of late recurrence and occult transmission by organ transplantation. Experimental data underscores the critical role of impaired angiogenesis and immune regulation as major mechanisms for maintenance of tumor dormancy. Finally, we examine evidence for the role of surgery in promoting escape from tumor dormancy at metastatic sites in cutaneous melanoma.

  5. Giant melanoacanthoma mimicking malignant melanoma

    Directory of Open Access Journals (Sweden)

    Vikas Shankar

    2011-01-01

    Full Text Available Melanoacanthoma denotes a rare variant of pigmented seborrheic keratosis. A 65-year-old male farmer had pigmented, verrucous, itchy, highly painful, progressively growing irregularly oval plaque on left side of lower back for the past five years. The indurated lesion, measuring maximum diameter 10 cm Χ 5 cm, had no discharge, bleeding, ulceration, or associated lymphadenopathy. Dermoscopy showed regular pigmentary network and cribiform pattern of ridges without any feature of malignant melanoma. Histopathology showed well-defined islands of basaloid cells interspersed with large and richly dendritic melanocytes. The lesion was totally excised followed by skin grafting. Our patient was unique in its massive size and clinical resemblance with malignant melanoma. The diagnosis was confirmed by dermoscopy and skin biopsy.

  6. Neutron capture therapy for melanoma

    International Nuclear Information System (INIS)

    The development of boron-containing compounds which localize selectively in tumor may require a tumor-by-tumor type of approach that exploits any metabolic pathways unique to the particular type of tumor. Melanin-producing melanomas actively transport and metabolize aromatic amino acids for use as precursors in the synthesis of the pigment melanin. It has been shown that the boron-containing amino acid analog p-borono-phenylalanine (BPA) is selectively accumulated in melanoma tissue, producing boron concentrations in tumor that are within the range estimated to be necessary for successful boron neutron capture therapy (BNCT). We report here the results of therapy experiments carried out at the Brookhaven Medical Research Reactor (BMRR). 21 refs., 5 figs., 3 tabs

  7. Snake venoms components with antitumor activity in murine melanoma cells; Componentes derivados de venenos de serpentes com acao antitumoral em celulas de melanoma murino

    Energy Technology Data Exchange (ETDEWEB)

    Queiroz, Rodrigo Guimaraes

    2012-07-01

    Despite the constant advances in the treatment of cancer, this disease remains one of the main causes of mortality worldwide. So, the development of new treatment modalities is imperative. Snake venom causes a variety of biological effects because they constitute a complex mixture of substances as disintegrins, proteases (serine and metalo), phospholipases A2, L-amino acid oxidases and others. The goal of the present work is to evaluate a anti-tumor activity of some snake venoms fractions. There are several studies of components derived from snake venoms with this kind of activity. After fractionation of snake venoms of the families Viperidae and Elapidae, the fractions were assayed towards murine melanoma cell line B16-F10 and fibroblasts L929. The results showed that the fractions of venom of the snake Notechis ater niger had higher specificity and potential antitumor activity on B16-F10 cell line than the other studied venoms. Since the components of this venom are not explored yet coupled with the potential activity showed in this work, we decided to choose this venom to develop further studies. The cytotoxic fractions were evaluated to identify and characterize the components that showed antitumoral activity. Western blot assays and zymography suggests that these proteins do not belong to the class of metallo and serine proteinases. (author)

  8. Melanoma of the female urethra

    OpenAIRE

    Ramos, Juan A.; Ramos, Wilmer E.; Ramos, Claudia V.

    2011-01-01

    Melanoma is a malignant tumor that can affect any area of the anatomical economy. Its appearance in the female urethra is extremely rare, with approximately 121 cases in indexed literature since 1966. The subject to be described is an 86-year-old woman who seeks assessment for intermittent macroscopic hematuria with blood clots of 3 months progression. On physical examination, there are no suspicious lesions detected on the surface of the skin. On external genital examination, it is observed ...

  9. Malignant rectal melanoma. Case report.

    Science.gov (United States)

    Morlino, Andrea; La Torre, Giuseppe; Vitagliano, Giulia; Cammarota, Aldo

    2015-03-26

    Il Melanoma Anorettale è una malattia rara e aggressiva ed è il terzo tipo più comune di melanoma maligno dopo quello della cute e della retina. Il sintomo più comune è il sanguinamento rettale, che è spesso scambiato per sanguinamento associato a emorroidi. La diagnosi è molto difficile, e quella iniziale può essere corretta solo in circa 80% dei casi. Il caso clinico che proponiamo riguarda un uomo di 71 anni giunto alla nostra osservazione per dolore anale, tenesmo rettale, sanguinamento. L’eplorazione rettale ci ha mostrato una neofromazione dolorosa, di colorito brunastro nel canale anale. La colonscopia e la endoscopia hanno evidenziato la presenza di una grande massa stenotica interessante il canale anale ed il retto con un diametro di circa 90 mm. La biopsia è positiva per melanoma a cellule maligne pigmentate. La TAC ha mostrato un ispessimento della parete rettale e linfonodi nel tessuto adiposo, nel distretto otturatore bilaterale e metastasi polmonari bilaterali. Il dato di laboratorio del Ca 19-9 è nei livelli normali. Il paziente è stato sottoposto a resezione addomino-perineale con dissezione linfonodale. Non ci sono studi dimostranti che la resezione radicale del melanoma primario ano-rettale è associata ad un miglioramento del controllo locale e della sopravvivenza. I pazienti con malattia localizzata dovrebbero essere sottoposti a escissione locale ogniqualvolta ciò sia tecnicamente fattibile. Il ruolo predominante del trattamento chemio radioterapico preoperatorio è quello di ridurre le recidive locoregionale e della cavità pelvica, e per ottenere un più alto tasso di conservazione dell’apparato sfinteriale. Inoltre facilita la rimozione delle potenziali micrometastasi e riduce le metastasi a distanza.

  10. Perineural extension of facial melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Kalina, Peter [Mayo Clinic, Department of Radiology, Rochester, Minnesota (United States); Bevilacqua, Paula

    2005-05-01

    A 64-year-old man presented with a pigmented cutaneous lesion on the right side of his face along with right facial numbness. Histological examination revealed malignant melanoma. Magnetic resonance imaging (MRI) revealed perineural extension along the entire course of the maxillary division of the right trigeminal nerve. This is a rare but important manifestation of the spread of head and neck malignancy. (orig.)

  11. UVB: suscetibilidade no melanoma maligno UVB: susceptibility in malignant melanoma

    Directory of Open Access Journals (Sweden)

    Nilton Nasser

    2010-12-01

    Full Text Available FUNDAMENTOS: Está bem definido que a radiação ultravioleta provoca depleção imunológica na pele, permitindo o desenvolvimento de tumores cutâneos malignos. A maioria dos pacientes de cânceres da pele não melanomas são considerados UVB-suscetíveis. OBJETIVOS: Estudar a UVB-suscetibilidade nos pacientes com melanoma maligno e se este é um fator de risco para o desenvolvimento desse câncer. MÉTODOS: Foram selecionados 88 voluntários divididos em dois grupos: grupo-controle saudável (n=61 e grupo de portadores de melanoma (n=27, todos identificados de acordo com os critérios: tipo histológico, nível de invasão, fotótipos de pele, sexo e idade. A suscetibilidade à radiação ultravioleta B (UVB foi medida pela reação de hipersensibilidade ao contato com o difenciprone nos voluntários sensibilizados em áreas previamente irradiadas. RESULTADOS: A suscetibilidade à radiação UVB foi de 81,5% nos pacientes com melanoma maligno e de 31,2% no grupo-controle. O risco de um indivíduo desenvolver o melanoma maligno foi 9,7 vezes maior do que nos indivíduos UVB-resistentes. CONCLUSÕES: A UVB-suscetibilidade pode ser considerada um fator de risco importante para o desenvolvimento do melanoma maligno.BACKGROUND: It is well established that UV radiation provokes an immunological depletion in the skin, enabling the development of malignant cutaneous tumors. Most nonmelanoma skin cancer patients are considered to be UVB-susceptible. OBJECTIVE: To study the behavior of UVB- susceptibility in malignant melanoma (MM patients and whether this is a risk factor to the development of MM. METHODS: Eighty-eight volunteers were selected and divided into two groups: healthy control group (n = 61 and MM group (n = 27, which were identified according to the following clinical criteria: histopathological type, level of invasion, skin phototype, sex and age. Susceptibility to ultraviolet B (UVB radiation was measured by the onset of a contact

  12. Experimental stress analysis and fatigue tests of five 24-in. NPS ANSI Standard B16.9 tees

    International Nuclear Information System (INIS)

    Experimental stress analyses and low-cycle fatigue tests of five 24-in. nominal pipe size American National Standards Institute (ANSI) Standard B16.9 forged tees are documented in this report. The tees, designated as Oak Ridge National Laboratory tees T10, T11, T12, T13, and T16, were tested under subcontract at Combustion Engineering, Inc. in Chattanooga, Tennessee. Experimental stress analyses were conducted for 12 individual loadings on each tee. Each test model was instrumented with approx. 225, 1/8-in. three-gage, 450 strain rosettes on the inside and outside surfaces; and 6 linear variable differential transformers mounted on special nonflexible holding frames for measuring deflections and rotations of the pipe extensions. Following completion of the strain-gate tests, each tee was fatigue tested to failure with either a fully reversed displacement controlled in-plane bending moment on the branch or a cyclic internal pressure that ranged from a value slightly above zero to about 90% of the nominal yield pressure of the pipe extensions

  13. Choroidal Metastases From Cutaneous Melanoma.

    Science.gov (United States)

    Mercado, Carmel L; Toy, Brian C; Kistler, Henry B; Moshfeghi, Darius M

    2016-05-01

    A 92-year-old man presented with months of progressive blurry vision, worsening acutely in his right eye. He denied pain, diplopia, or photopsias. His history was significant for multiple myeloma, prostate cancer, and malignant melanoma of his right shoulder treated with local excision. He had local recurrence with hepatic metastasis of the melanoma treated with radiation and chemotherapy. On examination, his visual acuity was counting fingers in the right eye and 20/60 in the left eye. Amsler grid testing demonstrated metamorphopsia in the right eye. Fundus exam of the right and left eyes revealed multiple, elevated, pigmented choroidal lesions, with associated subretinal fluid in the right macula. This appearance is consistent with hematogenous metastasis of cutaneous malignant melanoma to the choroid and associated serous fluid-causing metamorphopsia. The patient was enrolled in a clinical trial combining plasmid IL-12 with pembrolizumab (Keytruda; Merck, Whitehouse Station, NJ). He passed away 2 months after initial presentation to our clinic. [Ophthalmic Surg Lasers Imaging Retina. 2016;47:497.]. PMID:27183558

  14. CARI III Inhibits Tumor Growth in a Melanoma-Bearing Mouse Model through Induction of G0/G1 Cell Cycle Arrest

    Directory of Open Access Journals (Sweden)

    Hye-Jin Park

    2014-09-01

    Full Text Available Mushroom-derived natural products have been used to prevent or treat cancer for millennia. In this study, we evaluated the anticancer effects of CARI (Cell Activation Research Institute III, which consists of a blend of mushroom mycelia from Phellinus linteus grown on germinated brown rice, Inonotus obliquus grown on germinated brown rice, Antrodia camphorata grown on germinated brown rice and Ganoderma lucidum. Here, we showed that CARI III exerted anti-cancer activity, which is comparable to Dox against melanoma in vivo. B16F10 cells were intraperitoneally injected into C57BL6 mice to develop solid intra-abdominal tumors. Three hundred milligrams of the CARI III/kg/day p.o. regimen reduced tumor weight, comparable to the doxorubicin (Dox-treated group. An increase in life span (ILS% = 50.88% was observed in the CARI III-administered group, compared to the tumor control group. CARI III demonstrates anti-proliferative activity against B16F10 melanoma cells through inducing G0/G1 cell cycle arrest. CARI III inhibits the expression of cyclin D1, CDK4 and CDK2 and induces p21. Therefore, CARI III could be a potential chemopreventive supplement to melanoma patients.

  15. CARI III inhibits tumor growth in a melanoma-bearing mouse model through induction of G0/G1 cell cycle arrest.

    Science.gov (United States)

    Park, Hye-Jin

    2014-01-01

    Mushroom-derived natural products have been used to prevent or treat cancer for millennia. In this study, we evaluated the anticancer effects of CARI (Cell Activation Research Institute) III, which consists of a blend of mushroom mycelia from Phellinus linteus grown on germinated brown rice, Inonotus obliquus grown on germinated brown rice, Antrodia camphorata grown on germinated brown rice and Ganoderma lucidum. Here, we showed that CARI III exerted anti-cancer activity, which is comparable to Dox against melanoma in vivo. B16F10 cells were intraperitoneally injected into C57BL6 mice to develop solid intra-abdominal tumors. Three hundred milligrams of the CARI III/kg/day p.o. regimen reduced tumor weight, comparable to the doxorubicin (Dox)-treated group. An increase in life span (ILS% = 50.88%) was observed in the CARI III-administered group, compared to the tumor control group. CARI III demonstrates anti-proliferative activity against B16F10 melanoma cells through inducing G0/G1 cell cycle arrest. CARI III inhibits the expression of cyclin D1, CDK4 and CDK2 and induces p21. Therefore, CARI III could be a potential chemopreventive supplement to melanoma patients. PMID:25221864

  16. Pancreas Transplantation

    OpenAIRE

    Han, Duck Jong; Sutherland, David ER

    2010-01-01

    Diabetes mellitus is generally treated with oral diabetic drugs and/or insulin. However, the morbidity and mortality associated with this condition increases over time, even in patients receiving intensive insulin treatment, and this is largely attributable to diabetic complications or the insulin therapy itself. Pancreas transplantation in humans was first conducted in 1966, since when there has been much debate regarding the legitimacy of this procedure. Technical refinements and the develo...

  17. Copper-62 labeled ReCCMSH peptide analogs for melanoma PET imaging.

    Science.gov (United States)

    Zhang, Xiuli; Yue, Zhiwei; Lu, Bao-Yuan; Vazquez-Flores, Gerson J; Yuen, Johnny; Figueroa, Said Daibes; Gallazzi, Fabio; Cutler, Cathy; Quinn, Thomas P; Lacy, Jeffrey L

    2012-10-01

    High-specific activity radiolabeled melanocortin peptide preparations are necessary for optimal melanoma imaging due to the relatively low number of melanocortin-1 receptors (MC1-Rs) per tumor cell. In this study, a one-step synthesis of 62Cu-labeled MC1-R targeting peptide Re(Arg11)CCMSH was developed, which yielded high specific activity radiolabeled peptide preparations that required no post-labeling purification. DOTA and NOTA conjugated Re(Arg11)CCMSH peptides were synthesized and examined for 62Cu radiolabeling and cell binding properties. Biodistribution and PET imaging studies were performed to assess the in vivo tumor targeting and imaging characteristics of the optimal radiolabeled peptide. Melanoma cell binding affinities for NOTA-, NOTA-GGG-, and NOTA-GSG- conjugated Re(Arg11)CCMSH were determined to be 1.3×10-9 M, 1.9×10-9 M and 6.0×10-9 M. The 62Cu radiolabeling efficiencies of DOTA- and NOTA- conjugated Re(Arg11)CCMSH analogs were 30% and > 98% after 2 min at 24° C, while 0.5 μg of NOTA-GGG-peptide could be labeled to > 95% with a maximum specific activity of 138 Ci/μmol. Tumor uptake of 62Cu- NOTA-GGG-Re(Arg11)CCMSH in B16/F1 melanoma bearing mice was 4.65±0.48% ID/g and 9.43±2.69% ID/g at 20 and 40 min post injection and was visualized by PET imaging. High specific activity 62Cu-NOTA-GGG-Re(Arg11)CCMSH was prepared in a one-step procedure at 24°C in 6 min. 62Cu-NOTA-GGG-Re(Arg11)CCMSH exhibited MC1-R selective binding and rapid tumor uptake in B16/F1 melanoma bearing mice that was confirmed by PET imaging studies. High specific activity 62Cu from a 62Zn/62Cu generator coupled with simple one step radiolabeling procedures makes 62Cu an attractive radionuclide for PET imaging of low-density receptor targets. PMID:22724422

  18. Histological evaluation of intratumoral myxoma virus treatment in an immunocompetent mouse model of melanoma

    Directory of Open Access Journals (Sweden)

    Doty RA

    2013-01-01

    Full Text Available Rosalinda A Doty,1 Jia Liu,2 Grant McFadden,2 Edward J Roy,3 Amy L MacNeill11Department of Pathobiology, University of Illinois, Urbana, IL, 2Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL, 3Department of Molecular and Integrative Physiology, University of Illinois, Urbana, IL, USAAbstract: Two recombinant myxoma viruses (MYXV expressing a fluorescent protein [MYXV-Tred] and MYXV-Tred encoding murine interleukin-15 [MYXV-IL15] were evaluated for therapeutic effects in an aggressive B16F10 melanoma model in immunocompetent mice. It was hypothesized that continuous expression of IL-15 within a tumor would recruit cytotoxic effector cells to induce an antitumor immune response and improve treatment efficacy. Weekly intratumoral injections were given to evaluate the effect of treatment on the median survival time of C57BL/6 mice bearing established B16F10 melanomas. Mice that received MYXV-Tred or MYXV-IL15 lived significantly longer than mice given treatment controls. Unexpectedly, the median survival time of MYXV-IL15-treated mice was similar to that of MYXV-treated mice. At 1, 2, and 4 days postinoculation, viral plaque assays detected replicating MYXV-Tred and MYXV-IL15 within treated tumors. At these time points in MYXV-IL15-treated tumors, IL-15 concentration, lymphocyte grades, and cluster of differentiation-3+ cell counts were significantly increased when compared to other treatment groups. However, viral titers, recombinant protein expression, and lymphocyte numbers within the tumors diminished rapidly at 7 days postinoculation. These data indicate that treatment with recombinant MYXV should be repeated at least every 4 days to maintain recombinant protein expression within a murine tumor. Additionally, neutrophilic inflammation was significantly increased in MYXV-Tred- and MYXV-IL15-treated tumors at early time points. It is speculated that neutrophilic inflammation induced by intratumoral

  19. The potential effect of patulin on mice bearing melanoma cells: an anti-tumour or carcinogenic effect?

    Science.gov (United States)

    Boussabbeh, Manel; Ben Salem, Intidhar; Rjiba-Touati, Karima; Bouyahya, Chedy; Neffati, Fadwa; Najjar, Mohamed Fadhel; Bacha, Hassen; Abid-Essefi, Salwa

    2016-05-01

    Mycotoxins are bioactive compounds that are noxious to human. Their effects on oncogenesis have been satisfactorily elucidated, and some of mycotoxins have been classified as carcinogenic to humans. Nevertheless, patulin (PAT) is considered by the International Agency of Research on Cancer as 'not carcinogenic to humans'. The present study was designed to understand the effect of this mycotoxin on melanoma cells (B16F10) by measuring cell proliferation and assessing the anti-tumour effect in vivo in Balb/c mice. Our results revealed that intraperitoneally administration of PAT for 20 days significantly induces tumour regression in B16F10 cell-implanted mice. This effect was evidenced by the activation of apoptosis which is supported by the increase in p53 and Bax expressions, the downregulation of the protein levels of Bcl2, and the increase in caspase-3 activity. Moreover, systemic toxicity analysis demonstrated that there is no potential toxicity following PAT treatment unlike untreated melanoma mice which suffer from anaemia, inflammation and liver dysfunction. Remarkably, this is the first published report demonstrating the therapeutic efficacy of PAT in vivo models. PMID:26619846

  20. Design and preclinical evaluation of melanoma targeting agents for internal radionuclide therapy

    Energy Technology Data Exchange (ETDEWEB)

    Chezal, J.M.; Papon, J.; Labarre, P.; Denoyer, D.; Bonnet-Duquennoy, M.; Miot-Noirault, E.; Bayle, M.; Chavignon, O.; Teulade, J.C.; Maublant, J.; Madelmont, J.C.; Moins, N. [Centre Jean Perrin, Inserm-Univ. d' Auvergne, UMR 484, 63 - Clermont Ferrand (France)

    2008-02-15

    Targeted internal radionuclide therapy would be an effective alternative to current therapies for disseminated melanoma treatment. N-(2-diethylaminoethyl)-4-iodo benzamide (B.Z.A.) and compounds of this series exhibit a specific affinity for melanoma tissue giving them a potent application for gamma imaging ({sup 123}I) or radionuclide therapy ({sup 131}I or {sup 125}I).With the aim of internal radionuclide therapy, a pharmaco-chemical study has been carried out in order to select new derivatives with a longer retention time in the tumor of melanoma bearing mice and suitable dosimetry. New molecules synthesized are B.Z.A. analogs differing by (1) the aliphatic side chain, (2) aromatic ring. After the design of compounds, the synthesis, and the labelling with {sup 125}I, a study of their biodistribution was performed in B 16 F0 melanoma bearing C.57 B.L.6 mice after i.v. injection. The radioactivity biodistribution was analysed using an A.M.B.I.S.4000 detector on whole body slices of mice obtained by cryo section. The radioactivity was quantified in different organs including tumor and expressed as percentage of injected dose/g of tissue (% I.D./g). Dosimetry parameters for a {sup 131}I utilization were extrapolated using the M.I.R.D. program. The first selected compound has been further evaluated on different models. S.I.M.S. imaging, metabolism and in vivo anti tumoral activity after {sup 131}I labelling were assessed. For number of the studied molecules, a tumor uptake was observed and at least four compounds exhibited an original pharmacokinetic profile: high, specific and durable tumour concentration with a rapid clearance from non-target organs. The tumour concentration after 72 h was increased up to 16- fold compared to B.Z.A. and in term of dosimetry, for a {sup 131}I labelling, the tumor absorbed dose was increased by more than 6-fold. Such profiles made these compounds promising for an application to internal radionuclide therapy. The first selected

  1. Loss of retrovirus production in JB/RH melanoma cells transfected with H-2Kb and TAP-1 genes.

    Science.gov (United States)

    Li, M; Xu, F; Muller, J; Huang, X; Hearing, V J; Gorelik, E

    1999-01-20

    JB/RH1 melanoma cells, as well as other melanomas of C57BL/6 mice (B16 and JB/MS), express a common melanoma-associated antigen (MAA) encoded by an ecotropic melanoma-associated retrovirus (MelARV). JB/RH1 cells do not express the H-2Kb molecules due to down-regulation of the H-2Kb and TAP-1 genes. When JB/RH1 cells were transfected with the H-2Kb and cotransfected with the TAP-1 gene, it resulted in the appearance of H-2Kb molecules and an increase in their immunogenicity, albeit they lost expression of retrovirus-encoded MAA recognized by MM2-9B6 mAb. Loss of MAA was found to result from a complete and stable elimination of ecotropic MelARV production in the H-2Kb/TAP-1-transfected JB/RH1 cells. Northern blot analysis showed no differences in ecotropic retroviral messages in MelARV-producing and -nonproducing melanoma cells, suggesting that loss of MelARV production was not due to down-regulation of MelARV transcription. Southern blot analysis revealed several rearrangements in the proviral DNA of H-2Kb-positive JB/RH1 melanoma cells. Sequence analysis of the ecotropic proviral DNA from these cells showed numerous nucleotide substitutions, some of which resulted in the appearance of a novel intraviral PstI restriction site and the loss of a HindIII restriction site in the pol region. PCR amplification of the proviral DNAs indicates that an ecotropic provirus found in the H-2Kb-positive cells is novel and does not preexist in the parental H-2Kb-negative melanoma cells. Conversely, the ecotropic provirus of the parental JB/RH1 cells was not amplifable from the H-2Kb-positive cells. Our data indicate that stable loss of retroviral production in the H-2Kb/TAP-1-transfected melanoma cells is probably due to the induction of recombination between a productive ecotropic MelARV and a defective nonecotropic provirus leading to the generation of a defective ecotropic provirus and the loss of MelARV production and expression of the retrovirus-encoded MAA.

  2. Liver transplant - series (image)

    Science.gov (United States)

    ... depending on the degree of illness. After liver transplantation, patients must take immunosuppressive medications for the rest of their lives to prevent immune rejection of the transplanted organ.

  3. Mistletoe in the treatment of malignant melanoma

    Directory of Open Access Journals (Sweden)

    Esin Sakallı Çetin

    2014-03-01

    Full Text Available Malignant melanoma is a malignant neoplasia drives from melanocytes. Malignant melanoma, the most causing death, is seen in the third place at skin cancer. Malignant melanoma shows intrinsic resistance to chemotherapeutic agents and variability in the course of the disease which are distinct features separating from other solid tumors. These features prevent the development and standardization of non-surgical treatment models of malignant melanoma. Although there is a large number of chemotherapeutic agents used in the treatment of metastatic malignant melanoma, it hasn’t been demonstrated the survival advantage of adjuvant treatment with chemotherapeutic agents. Because of the different clinical course of malignant melanoma, the disease is thought to be closely associated with immune system. Therefore, immunomodulatory therapy models were developed. Mistletoe stimulates the immune system by increasing the number and activity of dendritic cells, thus it has been shown to effect on tumor growth and metastasis of malignant melanoma patient. Outlined in this review are the recent developments in the understanding the role of mistletoe as a complementary therapy for malignant melanoma. J Clin Exp Invest 2014; 5 (1: 145-152

  4. Unusual thoracic manifestation of metastatic malignant melanoma

    Directory of Open Access Journals (Sweden)

    Mohan K

    2010-01-01

    Full Text Available Massive pleural effusion due to metastatic malignant melanoma is rare. We report a case of bilateral (massive on left side pleural effusion as a metastatic manifestation of cutaneous malignant melanoma. In our case, successful outcome of pleurodesis with vincristine is significant as this agent is rarely used.

  5. Update in genetic susceptibility in melanoma.

    Science.gov (United States)

    Potrony, Miriam; Badenas, Celia; Aguilera, Paula; Puig-Butille, Joan Anton; Carrera, Cristina; Malvehy, Josep; Puig, Susana

    2015-09-01

    Melanoma is the most deadly of the common skin cancers and its incidence is rapidly increasing. Approximately 10% of cases occur in a familial context. To date, cyclin-dependent kinase inhibitor 2A (CDKN2A), which was identified as the first melanoma susceptibility gene more than 20 years ago, is the main high-risk gene for melanoma. A few years later cyclin-dependent kinase 4 (CDK4) was also identified as a melanoma susceptibility gene. The technologic advances have allowed the identification of new genes involved in melanoma susceptibility: Breast cancer 1 (BRCA1) associated protein 1 (BAP1), CXC genes, telomerase reverse transcriptase (TERT), protection of telomeres 1 (POT1), ACD and TERF2IP, the latter four being involved in telomere maintenance. Furthermore variants in melanocortin 1 receptor (MC1R) and microphthalmia-associated transcription factor (MITF) give a moderately increased risk to develop melanoma. Melanoma genetic counseling is offered to families in order to better understand the disease and the genetic susceptibility of developing it. Genetic counseling often implies genetic testing, although patients can benefit from genetic counseling even when they do not fulfill the criteria for these tests. Genetic testing for melanoma predisposition mutations can be used in clinical practice under adequate selection criteria and giving a valid test interpretation and genetic counseling to the individual. PMID:26488006

  6. Serum-proteomics in melanoma patients

    International Nuclear Information System (INIS)

    The project Serum-proteomics in melanoma patients funded by 'Programma Oncologico Italia-USA' Oncoproteomica has the general aim to collect serum samples from melanoma patients and to analyze the expression profile of several cytokines, in order to identify whether significant differences are evident between patients and controls, or among different patients subgroups with different staging or therapy

  7. Angiogenic and Metastatic Determinants of Malignant Melanoma

    NARCIS (Netherlands)

    A. Mooppilmadham Das (Asha)

    2015-01-01

    markdownabstractCutaneous melanoma or malignant melanoma of the skin is a highly metastatic disease, with an increasing rate of incidence, poor prognosis and high resistance to therapeutic intervention. Although early diagnosis and surgical resection of the primary lesion could significantly improve

  8. Malignant melanoma at a scientific laboratory

    International Nuclear Information System (INIS)

    The general consensus of the seven reviewers is that occupational exposures at Lawrence Livermore National Laboratory have not been established as a causal factor for the observed excess of malignant melanoma. Several observations support the impression that some or all of the observed melanoma excess may be attributable to intense surveillance and enhanced detection of early stage melanoma lesions. Since the incidence of melanomas among Laboratory employees has not diminished, an early harvesting effect is unlikely. This suggests the distinct possibility that localized, in situ melanomas that would normally not be detected are being reported, and that in the absence of this enhanced detection, many of these early stage lesions would show little or no clinical progression. This phenomenon would explain the continued high incidence of melanomas in the absence of a physical or chemical inciting cause. A key point in this reasoning is the issue of the rate of growth of early stage melanomas, and this point remains a key question for study. Even if the observed excess cannot be explained by detection bias, the reviewers agree that the Austin and Reynolds' study does not make a convincing case for occupational factors being a cause of the high melanoma incidence. 6 refs

  9. Extending Bragg peak of heavy ion beam and melanoma cell inactivation measurement

    Institute of Scientific and Technical Information of China (English)

    LiQiang; WeiZeng-Quan; 等

    1998-01-01

    A rotating range modulator was designed and manufactured.which is applied to extend Bragg peak of heavy ion beam.Bragg curves of 75MeV/u 16O and 75MeV/u 12C ion beams through this range modulator were measured respectively and two evident spread-out Bragg peaks corresponding to the modulated beams above are shown.In addition,inactivation effect of the modulated 75MeV/u 16O ion beam at nine different penetration depths on melanoma cells(B16) was measured.Results indicate that lethal effects at the spread-out Bragg peak region are larger than at the plateau of the particle beam entrance.

  10. Depigmenting Effect of Winter Medicinal Mushroom Flammulna velutipes (Higher Basidiomycetes) on Melanoma Cells.

    Science.gov (United States)

    Nagasaka, Reiko; Ishikawa, Yuki; Inada, Takumi; Ohshima, Toshiaki

    2015-01-01

    In humans, skin pigmentation results from the biochemichal synthesis and accumulation of melanin. We previously revealed that edible winter mushroom Flammulina velutipes extract inhibited melanosis in commercially available edible crustacea. Therefore, the current study was conducted to find if edible mushrooms extracts are useful for the skin whitening and the underlying mechanisms in melanin biosynthesis. Tyrosinase is one of the key enzymes for melanin biosynthesis. This study demonstrated that melanin synthesis and tyrosinase activity were suppressed by the F. velutipes extracts in B16 murine melanoma cells through the slow expression of microphthalmia-associated transcription factor. These results suggested that the F. velutipes extract could contain an effective component in whitening cosmetics and an alternative therapeutic agent for treating hyperpigmentation. PMID:26349509

  11. Immunological tumor destruction in a murine melanoma model by targeted LTalpha independent of secondary lymphoid tissue

    DEFF Research Database (Denmark)

    Schrama, D.; Voigt, H.; Eggert, A.O.;

    2008-01-01

    of specific T-cell responses even in the absence of secondary lymphoid organs. In addition, this effect is accompanied by the initiation of tertiary lymphoid tissue at the tumor site in which B and T lymphocytes are compartmentalized in defined areas and which harbor expanded numbers of tumor specific T cells...... as demonstrated by in situ TRP-2/K(b) tetramer staining. Mechanistically, targeted LTalpha therapy seems to induce changes at the tumor site which allows a coordinated interaction of immune competent cells triggering the induction of tertiary lymphoid tissue. CONCLUSION: Thus, our data demonstrate that targeted......BACKGROUND: We previously demonstrated that targeting lymphotoxin alpha (LTalpha) to the tumor evokes its immunological destruction in a syngeneic B16 melanoma model. Since treatment was associated with the induction of peritumoral tertiary lymphoid tissue, we speculated that the induced immune...

  12. Rotationally-Resolved Infrared Spectroscopy of the νb{16} Band of 1,3,5-TRIOXANE

    Science.gov (United States)

    Gibson, Bradley M.; Koeppen, Nicole; McCall, Benjamin J.

    2015-06-01

    1,3,5-trioxane is the simplest cyclic form of polyoxymethylene (POM), a class of formaldehyde polymers that has been proposed as the origin of distributed formaldehyde formation in comet comae and a potential source of formaldehyde in prebiotic chemistry. Although claimed POM detections have since been proven to be inconclusive, laboratory simulations of cometary conditions have yielded trioxane and other POMs While the microwave spectrum of 1,3,5-trioxane has been studied extensively, 4-7.}, to date only one rotationally-resolved ro-vibrational spectrum has been published. Here, we present our studies of the νb{16} band of gas-phase trioxane centered at 1177 wn. Trioxane was entrained in a supersonic expansion of argon and characterized by continuous-wave cavity ringdown spectroscopy using an etalon-stabilized external-cavity quantum cascade laser. Rotationally resolved spectra were obtained with less than 15 MHz resolution. Cottin, H., Bénilan, Y., Gazeau, M-C., and Raulin, F. Origin of Cometary Extended Sources from Degradation of Refractory Organics on Grains: Polyoxymethylene as Formaldehyde Parent Molecule. Icarus 167 (2004), 397-416. Oka, T., Tsuchiya, K., Iwata, S., and Morino, Y. Microwave Spectrum of s-Trioxane. Bull. Chem. Soc. Jpn. 37 (1964), 4-7. Henninot, J-F., Bolvin, H., Demaison, J., and Lemoine, B. The Infrared Spectrum of Trioxane in a Supersonic Slit Jet. J. Mol. Spect. 152 (1992), 62-68. Gibson, B.M. and McCall, B.J., contribution TJ08, presented at the 69th International Symposium on Molecular Spectroscopy, Urbana, IL, USA, 2014.

  13. Novel approaches in melanoma prevention and therapy.

    Science.gov (United States)

    Grimaldi, Antonio M; Cassidy, Pamela B; Leachmann, Sancy; Ascierto, Paolo A

    2014-01-01

    The incidence of cutaneous melanoma has risen at a rate significantly higher than that for other malignancies. This increase persists despite efforts to educate the public about the dangers of excess exposure to UV radiation from both the sun and tanning beds. Melanoma affects a relatively younger population and is notorious for its propensity to metastasize and for its poor response to current therapeutic regimens. These factors make prevention an integral component to the goal of decreasing melanoma-related mortality. Transformation of melanocytes into malignant melanoma involves the interplay between genetic factors, UV exposure, and the tumor microenvironment. The roles of UV radiation in the etiology of melanoma are mediated by both direct damage of DNA through formation of photoproducts and production of reactive oxygen species (ROS). Many of the promising antioxidant agents under development for the prevention of melanoma are derived from foodstuffs. B-Raf is a member of the Raf kinase family of serine/threonine-specific protein kinases that plays a role in regulating the MAP kinase/ERKs signaling pathway. About 50 % of melanomas harbor activating BRAF mutations. BRAF mutations are found in 59 % of the melanomas arising in skin with intermittent sun exposure, such as trunk and arms, as compared with only 23 % of the acral melanomas, 11 % of mucosal melanomas, and 0 % of uveal melanomas. Two new agents, ipilimumab and vemurafenib, have been shown to improve outcome of advanced melanoma as presented at the plenary session of the 2011 annual meeting of the American Society of Clinical Oncology. Vemurafenib is the first personalized compound which demonstrated an improvement in progression-free survival (PFS) and overall survival (OS) in metastatic melanoma harboring the BRAFV600 mutation and represents the first drug of a class that exerts its anti-proliferative activity through inhibition of a highly specific molecular target. GSK2118436 (dabrafenib), the

  14. Targeting melanoma growth and viability reveals dualistic functionality of the phosphonothionate analogue of carba cyclic phosphatidic acid

    Directory of Open Access Journals (Sweden)

    Prestwich Glenn D

    2010-06-01

    Full Text Available Abstract Background Although the incidence of melanoma in the U.S. is rising faster than any other cancer, the FDA-approved chemotherapies lack efficacy for advanced disease, which results in poor overall survival. Lysophosphatidic acid (LPA, autotaxin (ATX, the enzyme that produces LPA, and the LPA receptors represent an emerging group of therapeutic targets in cancer, although it is not known which of these is most effective. Results Herein we demonstrate that thio-ccPA 18:1, a stabilized phosphonothionate analogue of carba cyclic phosphatidic acid, ATX inhibitor and LPA1/3 receptor antagonist, induced a marked reduction in the viability of B16F10 metastatic melanoma cells compared with PBS-treated control by 80-100%. Exogenous LPA 18:1 or D-sn-1-O-oleoyl-2-O-methylglyceryl-3-phosphothioate did not reverse the effect of thio-ccPA 18:1. The reduction in viability mediated by thio-ccPA 18:1 was also observed in A375 and MeWo melanoma cell lines, suggesting that the effects are generalizable. Interestingly, siRNA to LPA3 (siLPA3 but not other LPA receptors recapitulated the effects of thio-ccPA 18:1 on viability, suggesting that inhibition of the LPA3 receptor is an important dualistic function of the compound. In addition, siLPA3 reduced proliferation, plasma membrane integrity and altered morphology of A375 cells. Another experimental compound designed to antagonize the LPA1/3 receptors significantly reduced viability in MeWo cells, which predominantly express the LPA3 receptor. Conclusions Thus the ability of thio-ccPA 18:1 to inhibit the LPA3 receptor and ATX are key to its molecular mechanism, particularly in melanoma cells that predominantly express the LPA3 receptor. These observations necessitate further exploration and exploitation of these targets in melanoma.

  15. Raspberry pulp polysaccharides inhibit tumor growth via immunopotentiation and enhance docetaxel chemotherapy against malignant melanoma in vivo.

    Science.gov (United States)

    Yang, Yong-Jing; Xu, Han-Mei; Suo, You-Rui

    2015-09-01

    It has been reported previously that the systemic efficacy of chemotherapeutic agents is substantially restricted for some cancer types, including malignant melanoma. Therefore, the development of more effective treatment modalities remains a critical, albeit elusive, goal in anticancer therapy. The study presented here evaluates the antitumor activity of raspberry pulp polysaccharides (RPPs) against malignant melanoma using a murine tumor-bearing model. Furthermore, the underlying mechanism of this antitumor activity has also been investigated. The results show that while RPP exhibits no direct cytotoxic effect on HT-29, MGC-803, HeLa, Bel-7402, L02 and B16F10 cells in vitro, it does demonstrate a dose-dependent growth inhibition of melanoma in vivo with an inhibition ratio of 59.95% at a dose of 400 mg kg(-1). Besides this, the body weight and spleen index in tumor-bearing mice have also been improved in RPP-treated groups. RPP is also found to induce splenocyte proliferation and is able to upregulate the activity of immune-related enzymes, including acid phosphatase (ACP), alkaline phosphatase (AKP), lactate dehydrogenase (LDH) and superoxide dismutase (SOD) in the spleen of tumor-bearing mice. The levels of tumor necrosis factor α (TNF-α), interferon γ (IFN-γ) and interleukin 2 (IL-2) in the serum of tumor-bearing mice show to be effectively increased upon RPP treatment. Histopathological analyses show that RPP induces tumor tissue necrosis by increasing inflammatory cell infiltration and causes no lesions to liver and kidney tissues. Remarkably, RPP further enhances the antitumor effect of the chemotherapeutic drug docetaxel and alleviates docetaxel-induced liver and kidney lesions in tumor-bearing mice. These findings indicate that RPP exhibits antitumor activity in vivo against malignant melanoma, partly by enhancing the cellular immune response of the host organism. In summary, RPP features critical properties to potentially find use as an

  16. Melanoma Surveillance in the US: Collecting Melanoma Data

    Centers for Disease Control (CDC) Podcasts

    2011-10-19

    This podcast accompanies the publication of a series of articles on melanoma surveillance in the United States, available in the November supplement edition of the Journal of the American Academy of Dermatology. Dr. Suephy Chen, a dermatologist from Emory University, discusses why the articles are important, as well as the need to increase dermatologists’ awareness of cancer registries and reporting requirements.  Created: 10/19/2011 by National Center for Chronic Disease Prevention and Health Promotion (NCCDPHP).   Date Released: 10/19/2011.

  17. Alpha particles for treatment of disseminated melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Link, E.M. [London Univ. (United Kingdom)

    2010-11-15

    Invading melanoma spreads to local and unpredictable distant location at the early stages of its development. It is justifiable, therefore to classify the disease as a systemic disorder. This requires a systemic treatment that reaches all melanoma cells irrespective of whether they are singly dispersed and in circulation or already forming solid tumours of various sizes. Targeted radiotherapy affects directly and selectively cancer cells provided an appropriate radionuclide and its carrier are chosen. Melanoma is a pigmented tumour. Methylene blue (MTB) accumulates selectively in melanoma cells due to its exceptionally high affinity to melanin. MTB serves, therefore, as a carrier for radionuclides. {sup 211}At-MTB has proved to be particularly effective in treating disseminated melanoma when administered systemically and, at the same time, non-toxic to normal non-pigmented and pigmented organs. (author)

  18. Alpha particles for treatment of disseminated melanoma

    Energy Technology Data Exchange (ETDEWEB)

    Link, E.M. [London University (United Kingdom)

    2010-07-01

    Invading melanoma spreads to local and unpredictable distant location at the early stages of its development. It is justifiable, therefore, to classify the disease as a systemic disorder. This requires a systemic treatment that reaches all melanoma cells irrespective of whether they are singly dispersed and in circulation or already forming solid tumours of various sizes. Targeted radiotherapy affects directly and selectively cancer cells provided an appropriate radionuclide and its carrier are chosen. Melanoma is a pigmented tumour. Methylene blue (MTB)) accumulates selectively in melanoma cells due to its exceptionally high affinity to melanin. MTB serves, therefore, as a carrier for radionuclides. {sup 211}At-MTB has proved to be particularly effective in treating disseminated melanoma when administered systemically and, at the same time, non-toxic to normal non-pigmented and pigmented organs. (authors)

  19. The risk of transmitting cutaneous malignancy through skin transplantation: a literature-based risk assessment.

    Science.gov (United States)

    Bosma, Sarah; Van Wijk, Marja J; Richters, Cornelia D; Beele, Hilde

    2015-12-01

    According to the European Union Tissues and Cells Directives donation of tissue is contraindicated in the presence of or a previous history of malignant disease, with the exception of cutaneous basal cell carcinoma. Skin cancer is the most common cancer. Due to ultraviolet light exposure and increasing life expectancy an increasing prevalence of malignant or premalignant skin lesions is observed, which may result in a decline of the availability of skin for transplantation. A risk assessment based on published studies and expert opinion was performed in order to investigate the risk of transmitting malignant or premalignant skin lesions through tissue transplantation, and more particular through skin transplantation. The scarcity of data concerning cancer transmission in tissue transplantation was challenging. Circumstantial evidence, available for organ transplantation, was used to develop the following policy proposal for skin transplantation and cutaneous tumours. Malignant melanoma is an absolute contraindication for the donation of skin and also of other tissues, whereas, non-lesional skin and other tissues of a donor with non-melanoma skin cancer (basal cell and squamous cell carcinoma) or with a premalignant skin lesion can be considered for transplantation. The above mentioned protocol proposal might serve as a prototype for analogous protocols for non-cutaneous malignancies. PMID:25631853

  20. Cytotoxic Activity and Antiproliferative Effects of Crude Skin Secretion from Physalaemus nattereri (Anura: Leptodactylidae on in vitro Melanoma Cells

    Directory of Open Access Journals (Sweden)

    Andréa Cruz e Carvalho

    2015-10-01

    Full Text Available Anuran secretions are rich sources of bioactive molecules, including antimicrobial and antitumoral compounds. The aims of this study were to investigate the therapeutic potential of Physalaemus nattereri skin secretion against skin cancer cells, and to assess its cytotoxic action mechanisms on the murine melanoma cell line B16F10. Our results demonstrated that the crude secretion reduced the viability of B16F10 cells, causing changes in cell morphology (e.g., round shape and structure shrinkage, reduction in mitochondrial membrane potential, increase in phosphatidylserine exposure, and cell cycle arrest in S-phase. Together, these changes suggest that tumor cells die by apoptosis. This skin secretion was also subjected to chromatographic fractioning using RP-HPLC, and eluted fractions were assayed for antiproliferative and antibacterial activities. Three active fractions showed molecular mass components in a range compatible with peptides. Although the specific mechanisms causing the reduced cell viability and cytotoxicity after the treatment with crude secretion are still unknown, it may be considered that molecules, such as the peptides found in the secretion, are effective against B16F10 tumor cells. Considering the growing need for new anticancer drugs, data presented in this study strongly reinforce the validity of P. nattereri crude secretion as a rich source of new anticancer molecules.